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Hung YW, Ouyang C, Ping X, Qi Y, Wang YC, Kung HJ, Ann DK. Extracellular arginine availability modulates eIF2α O-GlcNAcylation and heme oxygenase 1 translation for cellular homeostasis. Journal of biomedical science 2023 30(1) 37217939
Abstract:
Nutrient limitations often lead to metabolic stress during cancer initiation and progression. To combat this stress, the enzyme heme oxygenase 1 (HMOX1, commonly known as HO-1) is thought to play a key role as an antioxidant. However, there is a discrepancy between the level of HO-1 mRNA and its protein, particularly in cells under stress. O-linked β-N-acetylglucosamine (O-GlcNAc) modification of proteins (O-GlcNAcylation) is a recently discovered cellular signaling mechanism that rivals phosphorylation in many proteins, including eukaryote translation initiation factors (eIFs). The mechanism by which eIF2α O-GlcNAcylation regulates translation of HO-1 during extracellular arginine shortage (ArgS) remains unclear.
O-GlcNAc proteins:
TVBY1, TX13D, CC195, SPEM3, TVB42, FR1L5, HFM1, CCD66, SBNO1, Z804B, LRRD1, XIRP2, POTEF, PSD1, BDP1, NKX26, SWAHB, F181B, SMCO2, MGT4D, CAN8, EPOP, SMHD1, AKD1B, T200C, AQP73, AXA2L, NTM2B, ZAR1L, AN36A, NEU1B, GG6L6, B2R853, KDM4E, HNRC2, ERVV1, HNRC3, TEX22, TM232, MEIOS, CK098, SO1B7, TEX46, KIF2A, BACH, MYO1C, AP3B1, IKBE, HIP1, TULP1, WASL, PDE2A, SAP18, DNM1L, IFRD1, ZN593, FAAH1, PESC, SDCB1, MPP10, NOP56, DDX3X, CCN1, PLPP3, NCKP5, AGRB1, AP3D1, IDLC, CHD1, CHD2, PDCD5, ANM5, H17B6, FPGT, TCRG1, KIF3C, GEMI2, ML12B, CLGN, MYPT1, S27A2, XPO1, AT2A1, PLXB2, SR140, PER2, GRRE1, MCF2L, NACAD, ATS3, ANR28, VILL, LAMA5, MATN3, ATX7, INP4B, TRI18, GPR37, RPA34, MAGB3, PER1, KDM6A, DHX15, TTI1, PRP4, SERA, ADA12, IRAK2, ZW10, SNUT1, SPIT2, SRGP3, VIP2, PRPF3, GRID2, TGON2, LAT, SYT7, PLRG1, BUB3, PLPP2, PI15, ACTN4, SCO2, IDH3B, CALU, AGRB3, MED14, FGF17, PRKN, JIP4, KIF5C, ZC3H1, KATIP, OPA1, KIF1B, NMD3B, PEPL, PLXC1, ADCY9, HNRPQ, TPST1, RNBP6, CCNT1, FAIM3, ANM3, HCN1, RT14, HNRC1, H2B1K, PQBP1, PRAF2, DKC1, IF2P, TRI13, DNJA2, BRD4, NBN, IGSF3, DEN4B, ADA23, MFA3L, CLAP2, OBSL1, BRE1B, RFIP3, KDM4A, DJC13, ANR17, ZPR1, NIPS2, HMMR, ZN217, SC22B, PR40A, DPOLQ, ZN254, PSIP1, ERLN1, ECI2, KHDR3, TADA3, SF3B1, MED6, FOXH1, SPB7, U520, UTP20, TCEA3, SPI2, KLF7, MMP23, RASF9, CCNK, PRAF3, DYSF, SPF27, ATP5H, TRIO, RL1D1, NEBL, OCTN2, PDE9A, SRP72, B3GA3, UBP1, SLIT2, TOM70, MICA2, NFASC, FARP2, ERLN2, PRP6, GLSK, RNF37, WDR47, AP2A2, CE152, HEXI1, UBE4B, CSPG5, LETM1, LC7L3, KIF4A, GOSR1, KAT7, MRP6, TNKS1, KCNK5, VAPB, IPO7, M3K6, UTS2, CD2B2, FMNL1, SGPL1, G6PE, SC24A, ACSL3, DUS4L, ZBT11, CPSF4, K2C75, HERC2, SNP29, AP2A1, WIZ, STAU1, BAG2, ONEC2, TOP3B, APCL, PX11B, PAK4, LDHA, NB5R3, F13A, MOS, PGK1, TRYP, OVAL, IGHG1, IGHG2, IGHG3, IGHG4, HLAH, HBE, LMNA, ALBU, VTDB, TRFE, FRIH, MIS, VTNC, TDT, ALDOA, ANXA1, APOB, OAT, K2C1, SEMG1, TBB4A, G3P, NTRK1, P53, HSPB1, RPN1, RPN2, B3A2, AT1A1, CP17A, ITB2, HMGN1, CFAI, PCCA, IF2A, HMGN2, RLA1, RLA2, RLA0, AP2A, INSR, AGAL, RB, CDK1, ATPB, ENOA, NPM, TPM3, H2B1J, LDHB, H10, CATD, ANXA2, CO8A, TBB5, PROF1, HS90A, HNRPC, TPM2, GLI1, ACM2, CEL2A, HS90B, SRPRA, ODPA, CO4A2, MET, RU17, VIME, RS17, K2C7, ANXA5, K1C16, RSSA, GSTP1, RU1C, VILI, HMGB1, TPM1, WNT2, ROA1, C1S, PARP1, H2BFM, ATX1L, MYZAP, ZCC18, POTEI, POTEJ, SP9, UBB, UBC, MPC1L, ST1A4, HS71B, C1C1L, IGG1, D130B, M14OS, TBA3D, PR20G, TZAP, ACRO, H14, ODP2, THIO, PPGB, CH60, THA, BIP, HSP7C, ODPB, PYGB, PYGM, BCR, SPTB1, TOP1, PYC, DMD, C1TC, PABP1, K2C3, CO6A2, SKI, FETUA, ACTN1, MYH7, XRCC6, XRCC5, EF2, K1C10, K1C13, K2C5, TNNT1, COBA2, SUIS, KPYM, ENPL, CCNB1, HNRPL, MYOD1, EZRI, AREG, NQO1, RS2, CD44, H15, H13, H12, YBOX3, ZNF18, ZN182, ZNF43, HSP76, HMGA1, ZNF8, KPCA, CXA1, UBF1, DESM, NEUM, CR1, IBP3, TCPA, ITB5, RL7, SON, RL17, SDC1, K2C4, NUCL, E2AK2, CSK22, NCF2, FST, IF2B, ANXA7, CD33, BTF3, TNR1B, RAB3B, CAN3, ICAL, CO5A1, PHOS, CD11B, CSRP1, FLNA, PERL, TENX, PUR6, UBA1, H1T, ROA2, SAV, FBLN1, TCEA1, ITA6, SFPQ, PPIB, RS3, PTPRE, H2B1O, COF1, IF4B, CPT2, CCND1, RAE1, KPCL, THIL, RPB1, LAMA1, DNJB1, ATPA, DDX6, CTNA2, DNMT1, U2AF2, RL13, S10A4, HMGB2, PTBP1, HGFL, STOM, PON1, 1433T, RL10, CALR, MAP4, CALX, PSA5, ABCD3, LYOX, HXD10, PTPRM, PTN4, T2EB, EPHA2, EPHA8, TKT, LMOD1, EF1D, RL12, PDIA3, 2AAA, MPIP3, BKRB2, NKTR, KPYR, SDHA, HXA10, HXA9, GBRA5, DNJA1, AKT1, HNRH1, 1433B, S10AB, P5CR1, IFNA8, DSG3, RL9, KINH, CP2CI, H2B1B, EVI2B, HS71L, HSP74, PHB1, MERL, RADI, RL22, T2FA, SCN1A, K1C9, FBN1, FBN2, IRS1, MYH9, MYH10, COPB2, SOAT1, FUS, DEK, CYLC1, MYH11, AHR, K22E, ACHA7, RL4, LONM, ACV1B, SRP14, ZEB1, TAGL2, BRCA1, VATA, GRP75, RS19, RL3, OST48, FEN1, TXLNA, TCPZ, ARL1, UBP8, RFC3, ECHA, IF2G, PERI, BUD31, NAA10, KDM5C, GARS, SYIC, NBL1, LAP2A, LAP2B, EPS15, RS27, LPPRC, RL35, PF2R, MUC18, MATR3, GPDM, RANG, NAMPT, EFTS, KI67, RAGP1, ATRX, IF2M, BAG6, RL27A, RL5, RL21, RS9, RS5, RS10, MAP1B, IQGA1, IF1AX, OMP, RL29, ATPO, TCPE, K2C6C, KC1A, MASP1, HSP77, TCPG, EFTU, SRP09, SYAC, SYCC, SYSC, YLPM1, TMEDA, RBM25, TSC2, GUAA, ENTP1, MEOX1, ST1A1, ST1A2, GATM, SERPH, KNTC1, CDK9, R144A, RL14, TCPQ, TCPD, FXR1, RAB7A, ZNF83, SSRD, AL3A2, DHB4, PSMD7, CCL11, ZNF41, ROA3, MYOM1, HNRPM, IMA1, IMA5, MP2K6, HNRPF, MSH6, HMGA2, CRIP2, COPB, COPA, CRBB1, SMTN, MOT1, IST1, SC24C, TCP4, SYRC, ATX3, TERF1, SYYC, HSP72, ATNG, EPHA4, GYS2, DRG2, XPO2, MFAP1, PAR2, BRPF1, NP1L1, ANM2, PSA, EIF3B, SYMC, HDAC4, IF6, RP1, NRG3, H2BFS, UBS3A, WDR4, SNX16, SELB, SESN2, H2B1D, NAL11, DJB13, TPIS, MYPR, MYL6, ACTB, IF4A1, RS20, S10AA, CDC42, DEST, SRP54, RAB2A, RAB14, ZC3H6, ACTZ, ARF3, ABCE1, RS3A, RL26, RL15, RL27, RL37A, DCAF7, HNRPK, RRAS2, RS7, RS8, RS15A, RS16, 1433E, RS14, RS18, RS29, RS13, RS11, SMD1, SMD2, ARF6, RL7A, RS4X, ACTA, RL23A, RS6, VISL1, H4, H2B1C, RAB1A, RAN, RL23, RS15, RS24, RS25, RS26, RS30, RL39, RL31, RL10A, RL32, RL11, RL8, PPIA, RS27A, RL40, TRA2B, AP2B1, 1433Z, 2ABA, RL38, RACK1, ACTG, ACTH, YBOX1, CSK2B, TPM4, ACTC, EF1A1, ACTS, TBA1B, TBA4A, TBB4B, CSK21, HBG1, HBG2, NOP14, RPP30, PHC1, S35A1, PRKDC, M3K9, SSBP2, RT22, RT11, RT09, ATL3, RL24, RL19, SRSF3, MUC5A, TFAM, CLH1, NFKB2, HNRPU, MYPC1, MDM2, U2AF1, AMPD2, DIAC, CAP1, K22O, CAC1D, EXOSX, PFKAP, EWS, CO4A3, AK17A, CENPE, MMSA, BDH, DSG1, H11, GHRHR, M3K10, PLOD1, RL6, PO3F1, CAV1, KMT2A, TAP2, ZN117, IF4G1, SRS11, EF1A2, CALD1, AT5G2, PTN11, GABP1, ZN33A, FMR1, PRDX1, RL18, C1QBP, CKAP4, PDE4B, KHDR1, SLC31, SOS2, SRSF1, ACTN3, TLE5, SRSF4, DHX9, GOGA2, DSC1, SSRP1, CAMP2, NSUN2, AHNK, DIK1C, LR74A, LMOD3, CF210, GALT2, AP1B1, HMGX3, NU160, TBL3, TF3C1, ASPH, TROP, BPTF, CNTN1, SFSWA, CHD3, SF3A3, NMDE1, TP53B, ILF3, TIAM1, AKAP6, FLII, TRDN, ACACA, ZBT17, REST, TADBP, ROA0, CBX3, STK3, CBLB, DNJC3, M3K1, SRSF6, I15RA, G3BP1, ATM, AP3B2, PPIG, TCOF, SF3B2, GOGA4, ROCK1, TOP3A, SQSTM, MED21, MTX1, TBB3, PRP4K, ATR, KCC2D, TDG, SNW1, DGKZ, CUL1, S12A1, MOGS, NACA, THOC5, NOG2, PTK6, TBB2A, CAC1C, CKAP5, CO4A6, LRP8, PDE1C, BOP1, SEPT6, MORC3, EFR3A, UBP2L, SIM2, DYHC1, ELOA1, TRI25, MYPC2, FRG1, CAPR1, RBM39, SCN5A, PDIA5, HS902, ZN266, ZN235, SMC1A, RRP1B, NCOA6, GANAB, 2A5D, MVP, LTBP1, GOGB1, GRM7, CHD4, LASP1, RB39A, IMB1, PSME4, CUL7, NAA25, RTP5, MO4L2, SEPT2, SUZ12, U5S1, BRD3, EEA1, PDIA6, PCOC1, PDK2, PLEC, PCM1, NONO, RASA2, RNPS1, PCBP1, PCBP2, SF3B3, RYR3, CNN3, SF3B4, RBMS2, SF3A1, SIX1, MED22, TRAM1, SF01, TRIPB, OASL, MED1, TRIP6, TWST1, PI5PA, CEBPE, AAAT, TOM34, NCOA1, GRK1, SCN9A, ZYX, SEPT7, UAP1, GBRA6, CSRP2, CDC37, IL17, DPYL2, M3K11, SRSF7, CPSF6, AT2B3, H2A2C, H2B2E, HCDH, FA98C, TIGD6, NKPD1, KNOP1, ESRG, PDS5A, MOONR, BLTP1, KIF7, TSR1, C19L2, CO3, PRTG, GAB4, ZN750, P3H1, RA1L2, MA7D1, CHD9, TKFC, PTHB1, RABL6, TBB8, CRFM7, GLYR1, C2CD3, LARP7, TB10B, A20A4, GRAP1, MIDN, PKDCC, CN177, CEP55, KLH22, PLA1A, SGO2, ACTBL, BRDT, H90B4, H90B3, H90B2, HS905, ODFP2, ESCO1, SH319, AN36C, PPM1J, FGD3, RRP12, CF157, ZN484, GNAS1, GASP1, EOGT, BRM1L, H2B2F, PPM1L, TM201, CA167, TT39A, HP1B3, CE170, FREM2, CE85L, SZT2, AXDN1, EYS, ZC3HD, GPTC4, RBM20, UBR4, GNTK, ER6L2, HENMT, TTC22, FRY, B2L15, KAD9, M3K21, DDI2, MROH9, EFCB6, A20A1, CROCC, CL060, OBSCN, FA78B, EF1A3, PR38B, BRE1A, SMBT2, SPO16, TASO2, MALR1, ECM29, SHE, NAA35, TM268, RES18, K2C79, KFA, WSCD1, F91A1, CP135, AR6P4, MROH7, UTP25, RHG17, Z518A, SCAR3, KDM4D, IQEC1, XAF1, OVOS1, CAPR2, IQCE, ZN770, F166A, NIPBL, LIN54, TET2, TEN4, OGFD2, TM214, CAVN1, CDC73, ARMX5, MED27, ANM9, PRP8, PLBL1, NFRKB, ALKB5, TRI40, TBA3E, PONL1, TTC33, MYO7B, RICTR, BRWD3, POTEE, POTEA, CC171, S4A10, FIP1, PRS55, LRSM1, SBSN, CLM9, LRIG3, RSPO2, TMC5, RFIP1, CD109, HSDL2, B3GN6, CCNI2, M3K15, LN28B, NFXL1, DYH12, RN207, ITPI1, CRACD, YC006, PLPL7, FGD6, PHLP2, ANKS3, YS043, NEK10, KRA55, KRA52, VP13C, UBP34, RAPH1, UBP51, UBP45, UBP50, IKIP, MD13L, TBA1A, B4GN4, ATS13, HECW1, BT2A1, SND1, DDX46, TM10C, RSRC2, DET1, H2A3, FAKD5, NLRC3, SSX6, MOT9, OVCH1, PLPR4, PCLI1, NRK, MON2, K1C28, K1C27, K1C25, CSMD3, NUFP2, SZRD1, NPHP3, TT21B, GDF7, KI21A, CFA91, ZN438, HDGR2, SDK1, SPRE1, SRGP1, RBBP6, NECA2, SH3R1, SEN54, K2C1B, CNTRL, CTU1, RM55, VP13B, AGRE4, GLRX5, ZN605, DAAM2, K1328, PATL1, DDX53, TRPT1, LYRIC, AKA28, KTN1, GTD2A, ABCAD, EST1A, PHLB1, LUZP1, THOC4, IFTAP, LRRT4, ZCH18, ARHGP, MON1A, THOC6, AMGO1, AMGO3, AROS, CHSS1, DDX42, NIBA3, CEP57, DAAF5, LYPD6, STX12, DZIP1, HORN, KLOTB, WFDC8, GLT13, ZDH13, CRLF3, CRERF, ARMC8, K0825, ANKY2, AHNK2, DYH10, P3H2, SPNS2, VSTM4, WDFY1, PHF6, SULF2, CHERP, ANKH1, PHC2, OD3L1, F217A, DDIAS, RB12B, SP8, DDX60, RAVR1, SPB1, DRC7, EFC13, CKP2L, SRRM1, CCD13, ELOA2, CC105, ZN100, SMYD4, MICU2, CFA46, RN168, CHSS2, RTKN2, DOCK3, AGRF3, XRN1, ASPM, DSEL, RBIS, SPICE, TAGAP, NDUF2, VPS52, LRC47, K2C78, MYO1H, KLH34, H2B3B, ANR35, GAPT, UB2J2, AN36B, LAT4, EH1L1, PALS1, FNBP4, GUF1, RIBC1, NIN, ZN276, ZN561, SRBD1, NKAP, MACOI, PAR16, AADAT, IQCF1, EFNMT, KLDC1, PLD5, ANR31, CDYL2, AMER3, NEPR1, YJ012, MUT7, BANP, SR1IP, TOPZ1, CO053, PR38A, UBP38, UN13C, FANCB, GOLM1, GT251, DHB11, TM145, SERB1, DYHC2, CC020, GIMA8, TNR6A, ZN740, DIK2A, ZN738, ABCF1, DUS18, ZN548, EXPH5, KMT2C, ABHDB, MYRIP, KNL1, RDM1, O52E5, DDX55, MAFA, ZFP28, THOC2, KCNG3, TTC8, PI42C, TBC15, MIEAP, CIP2A, ZN507, CERK1, BICD2, CHD6, HELQ, CHD5, KCNG4, BRX1, CLM1, OTU7A, ATS16, DYH5, DQX1, TP4AP, WHAMM, UBS3B, AT8B4, SHRM3, WIPF2, F222B, ZC3HF, PPIL4, CCD12, HDAC7, PDC6I, CATIN, CO040, SFT2A, SNR27, HNRLL, UBCP1, CQ050, CTL1, CKAP2, SPIR2, ATX2L, ABCAA, SREK1, CRGN, SYNE2, MUC16, P66B, DNJC9, TITIN, DDX1, H1X, TM131, IP6K1, AP3S1, HS105, SEPT8, PRP16, AN32B, TFG, AP2C, STAM1, DPF3, REQU, RBP56, EVPL, DDX17, KIFA3, SEM4D, XPF, SMRD2, RAB1C, FUBP2, KCNB2, USP9X, UBP7, CUL5, VPP1, MED12, TATD2, H2B1H, FBXW7, YIPF5, UBE2F, RL36L, GTPB3, H2B1A, CCD47, FERM2, PLPL2, LRC59, PBIP1, TTC28, BORC7, SPSB1, ANKR9, CC127, DOC10, P5CR2, ROP1L, FAF2, ZN689, AP2M1, NTNG2, KCD12, PGBD4, MTBP, RMXL1, HOOK2, RBM33, DTBP1, TMA16, ALG14, CPNE2, KIF12, SIPA1, THOC1, RM48, DDX27, P121A, TONSL, PDLI5, PGAM5, COX14, AJUBA, A26L1, PAWR, VPS50, JPH4, NEUL4, HSH2D, ZN514, AGRG7, RPGR1, ZFR, NAR5, ALPK3, IFT74, PPR36, LRGUK, DNHD1, KLH15, ZN512, RIPR3, RN145, CFA44, ZDH15, PWP2A, DOCK7, ZN830, BROMI, SCLT1, CNT3B, ANR27, GP101, IPO9, LRC3B, PGRP2, INP4A, ADCYA, GBP4, ZN317, NED4L, SLIK1, NEK1, PP1RA, XPO6, H2A1A, MAGI1, SNX18, S26A8, MCCA, CTGE2, HMCN1, S22AC, RANB9, CCNL2, SMC6, ZN347, STRBP, SIN3A, RBM15, PNKP, SETB2, NIBA2, SCN2A, NELL2, PSB7, SEC62, CDC5L, STAC, AGAP2, GA2L1, DNJC2, PHB2, KIF2C, HCD2, CPNE1, EBP2, H2B1N, H2B1M, H2B1L, MYOC, SEM3C, GDF15, AKAP9, MICU1, TRIR, SLX1, MEP50, TBA1C, SELS, MBB1A, SYT3, NRIP2, FYCO1, GPTC1, RIOK1, LLPH, WDR83, S35A5, PIMRE, SOX7, WAC, MTA3, MCMBP, MED10, LRRC1, TBB6, HNRL1, DDX23, TBB2B, TMED9, RIOK2, ELOV1, RFIP5, ASPN, AP1M1, NUSAP, TDRD1, EMIL2, AN30B, SCAPE, RTF2, DUS16, MK67I, HOIL1, RT26, KRA41, SETD2, TBD2A, ACTBM, CNTP3, PC11X, PC11Y, PUS3, UACA, REN3B, CRNL1, EDEM3, UBXN6, API5, ZFHX2, CFA74, CE295, ASXL3, ZN407, LRC27, WDR12, IMUP, SENP6, CHST5, WN10A, EGLN1, PEG3, CHST6, NACA2, PAIP1, LSG1, KLC2, ZN541, INT2, DDX47, F234A, NUCKS, S6A17, ABCG5, EHD4, P2Y12, VPS11, TARA, SLK, TNKS2, RBP17, RM46, PININ, SNX25, MOD5, MTRFR, HRH4, NDST4, PTN23, SEM6B, MFSD1, PIGU, PLK3, FA83D, TRABD, EHD1, MROH8, PDZD7, TFB2M, DHX35, ZN576, BICC1, Z385D, CCD86, PRR36, RANB3, AT133, NRDE2, METL8, ZN430, CAAP1, NOL11, ZFAN3, F204A, VRTN, MARH7, SFXN1, SPE39, VP37B, UBP42, KANL2, LN28A, EAF6, ZN556, LIN7B, RENT2, XPO5, CP343, MKNK2, UBP29, PLRKT, SPC25, GPR35, SPON1, LST2, TANC2, EPG5, K1586, ZBT26, FBSL, ZN334, SPCS, MYO10, APMAP, ZCC17, VTA1, SYSM, SIX2, DCP1A, MED4, GP108, LUC7L, SCUB2, RM40, CASS4, RTN4, IFT46, RPR1B, ANLN, BIRC6, DDX21, SAR1A, ASH1L, ARHG4, ARHG3, MXRA5, SPTN5, SNRK, STRN4, TRYG1, SOSSC, F1142, LTBP3, KRT84, DIAP3, PDS5B, SMC4, MA2C1, RN146, MDN1, LYRM2, SPS2L, ABCF3, MRGBP, SEP11, MED17, FANCI, DDX18, KLH11, RPC5, SDA1, CS073, ARGL1, PK1IP, HPIP, CB042, SDHF2, NHP2, LYAR, MED29, CWC25, THUM1, SCN3A, TBA8, DYH9, HXC10, FKB11, CELR1, CDK12, FMN2, RTEL1, IL36B, SACS, E2AK3, PA2GE, MYOF, BICRA, NOP53, HACD3, KCMF1, RAI14, VAPA, SLAF8, SIRPG, HCN3, DYH2, RCC2, TEN3, RRBP1, CPSF2, SYLC, ANKY1, HOP2, IKBL1, COPG2, MYOTI, P2RX2, SPAST, TRMB, FA8A1, DBF4A, MO4L1, MALT1, DAXX, MARCO, ZBT47, CGBP1, GCP4, KDM5B, SYWM, HMGX4, NOL12, ARMX3, SWP70, SUN2, AFF4, MKRN1, NPCL1, CHRD1, SEPT9, PCS1N, NB5R1, S30BP, PUF60, SCNBA, DNAI1, BAZ2B, PHF11, VAX2, RABX5, TRM6, ZN391, APC7, STML2, VPS28, CCNL1, FBX9, MRT4, PITC1, GPTC8, PARP4, MYO15, TF3C4, ADA28, NUP50, ZN236, ZN346, HCN2, ZN214, NRX1A, ZN608, MRTFB, ATD2B, NEB1, SHRM4, TMCC3, MYO5B, TPX2, NP1L2, MAFF, NOTC3, PRP19, G3BP2, RHG26, CHIP, MAGD2, SSRG, ING4, DIM1, TTF2, L10K, LIMC1, AGAP1, TNR6B, ZC3H4, THS7A, RIMS2, SRRM2, CTND2, BAIP2, SCN8A, PPR3C, LAMP3, RTRAF, RUVB2, LZTS1, TN13B, SYFA, SIGL7, DRG1, NCKP1, TLR6, ATF5, DIP2C, DLGP4, FYV1, KDM2A, TPPC8, KLH20, S27A6, YBOX2, TR150, WBP11, NOP58, NOSIP, LC7L2, SF3B6, ORN, NOP16, TIM16, RT23, RTCB, HCN4, ZN330, RL36, CCDC9, HBS1L, DMXL1, ZAN, KDM3A, MFHA1, PLXD1, HECD4, WDR7, IRS2, HYOU1, OARD1, SP16H, PCDGH, HEY1, SRPRB, O51B2, CORIN, DMRT2, MRCKB, NOX1, RBM8A, ZN706, SNX9, SCNAA, HCFC2, PUS1, MYH4, SPIN1, COPG1, CFA20, EMIL1, PTC2, DC1L1, STYL1, OAS3, TLL2, ZNT1, MYO16, GAK19
Species: Sus scrofa, Homo sapiens, Bos taurus, Gallus gallus, Streptomyces avidinii
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Wong YK, Wang J, Lim TK, Lin Q, Yap CT, Shen HM. O-GlcNAcylation promotes fatty acid synthase activity under nutritional stress as a pro-survival mechanism in cancer cells. Proteomics 2022 22(9) 35083852
Abstract:
Protein O-GlcNAcylation is a specific form of protein glycosylation that targets a wide range of proteins with important functions. O-GlcNAcylation is known to be deregulated in cancer and has been linked to multiple aspects of cancer pathology. Despite its ubiquity and importance, the current understanding of the role of O-GlcNAcylation in the stress response remains limited. In this study, we performed a quantitative chemical proteomics-based open study of the O-GlcNAcome in HeLa cells, and identified 163 differentially-glycosylated proteins under starvation, involving multiple metabolic pathways. Among them, fatty acid metabolism was found to be targeted and subsequent analysis confirmed that fatty acid synthase (FASN) is O-GlcNAcylated. O-GlcNAcylation led to enhanced de novo fatty acid synthesis (FAS) activity, and fatty acids contributed to the cytoprotective effects of O-GlcNAcylation under starvation. Moreover, dual inhibition of O-GlcNAcylation and FASN displayed a strong synergistic effect in vitro in inducing cell death in cancer cells. Together, the results from this study provide novel insights into the role of O-GlcNAcylation in the nutritional stress response and suggest the potential of combining inhibition of O-GlcNAcylation and FAS in cancer therapy.
O-GlcNAc proteins:
RUXGL, ADAS, DX39A, MYO1C, IPO5, PESC, NOP56, DDX3X, SCD, MGST3, HNRDL, XPO1, SURF4, OGT1, PPM1G, MOT4, DHX15, CYB5B, SERA, HNRPR, BUB3, ACTN4, MYO1B, GANP, HNRPQ, NDUS7, MPU1, H2AY, FLNB, SC22B, SF3B1, U520, UTP20, NU155, ATP5H, RL1D1, MTA2, RTN3, VAPB, IPO7, ACSL3, BAG2, TOM40, LDHA, DHE3, AATM, PGK1, ASSY, LMNA, TFR1, ALDOA, K2C1, G3P, HSPB1, RPN1, AT1A1, ADT2, PCCA, RLA1, RLA0, LA, K1C18, K2C8, ATPB, ENOA, NPM, TPM3, LDHB, PDIA1, ANXA2, TBB5, TRY1, PROF1, SYEP, HS90A, HNRPC, DAF, 4F2, HS90B, ODPA, RU17, VIME, RS17, K2C7, GNAI3, RSSA, LEG1, ROA1, PARP1, PRS56, HS71B, ODP2, THIO, MGST1, CH60, BIP, HSP7C, GTR1, TOP2A, PYC, PABP1, PCNA, ADT3, IMDH2, KCRU, XRCC6, XRCC5, EF2, K1C10, K2C5, PDIA4, PLST, ETFA, MIF, KPYM, ENPL, HNRPL, PLAK, EZRI, NDKA, RS2, DESP, H13, NCPR, AT2A2, DDX5, TCPA, PTN1, ARF4, RL7, RL17, NUCL, GSTM3, FLNA, FBRL, PUR6, UBA1, ROA2, QCR2, SFPQ, PPIB, RS3, SAHH, COF1, MCM3, RS12, ATPA, U2AF2, RL13, S10A4, PTBP1, SYVC, EF1G, STOM, RL10, APEX1, PYR1, CALX, TKT, ERP29, PRDX6, PRDX5, PRDX3, RL12, PDIA3, CPSM, HNRH1, STIP1, L1CAM, PRDX2, P5CR1, DUT, MCM7, GLYM, HSP74, PHB1, RL22, MYH9, SOAT1, DEK, K22E, RL4, LONM, NUP62, GRP75, IF4A3, RL3, RL13A, ARL1, STAT3, MDHM, RFC3, ECHA, SYIC, LAP2A, LPPRC, MATR3, MSH2, GPDM, VDAC2, KI67, BAG6, RL27A, RL5, RS9, STT3A, CAPZB, SYQ, RL29, AT5G3, TCPE, RL34, FAS, TCPG, EFTU, ACADV, TMEDA, NU153, RBP2, CPT1A, SERPH, RL14, TCPQ, TCPD, FXR1, RAB5C, RAB7A, HCFC1, ROA3, 6PGD, HNRPM, IMA1, HNRPF, MSH6, TXTP, ACLY, COPA, MOT1, SYRC, KAD2, P5CS, XPO2, TERA, NP1L1, DSRAD, ATPK, TMM33, TPIS, MYL6, IF4A1, RS20, S10AA, RAP1B, RL15, RL37A, HNRPK, RS8, RS16, 1433E, RS14, RS23, RS11, RUXE, RL7A, RS4X, RS6, H4, RAB1A, RAN, RL23, RS25, RS26, RL10A, RL11, RL8, PPIA, RS27A, RSMN, RACK1, ACTG, UBC9, TBA1B, TBB4B, GTF2I, TCPB, PRKDC, RL24, ARF5, RL19, SRSF3, S25A3, CLH1, HNRPU, SPTB2, EXOSX, RL18A, RL6, IF4G1, K1C17, PRDX1, RL18, C1QBP, KHDR1, DHX9, NCBP1, AHNK, NU160, SF3A3, ILF3, ACACA, PRDX4, CBX3, TIF1B, SPTN1, HNRPD, SAFB2, TTL12, CAPR1, ITPR1, RRP1B, GANAB, LBR, GOGB1, IMB1, NUMA1, SUZ12, U5S1, RRS1, PDIA6, PLEC, TEBP, NONO, PCBP1, PCBP2, DHC24, SF3B3, SF3A1, TRAM1, ELAV1, AAAT, RBBP7, H31T, PDS5A, TSR1, IF2GL, RRP12, NU188, HP1B3, EF1A3, PPR18, PRP8, C1TM, DHX30, CAND1, MISP, SPB1, PELP1, RDH10, CCAR2, TXND5, STT3B, BRX1, PO210, GEMI5, RT27, HS105, GCN1, NU205, AKAP1, AN32B, RBP56, DDX17, FUBP2, TNPO1, UBP7, UTP4, LRC59, PGAM5, FUBP3, MBOA7, MCCA, WRIP1, UHRF1, POP1, HCD2, ROAA, TM9S2, TCPH, ANM1, H2B1L, RNZ2, MEP50, MBB1A, ESYT1, H2AJ, GNL3, HDHD5, GTPB4, API5, RPF2, SFXN1, RDH14, ABCB6, DDX21, MDN1, DCA13, ATD3A, DDX18, MIC19, TEX10, TECR, MYOF, THYN1, HACD3, RRBP1, ABC3B, RLP24, ACINU, OGDHL, COR1C, PRP19, SSRG, TRI33, EIF3L, RUVB1, VDAC3, PDIP2, NOP58, SF3B6, RTCB, RL36, LAS1L, SRPRB, COPG1, MTCH2, CEPT1, ZNT1
Species: Homo sapiens
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Fan Z, Li J, Liu T, Zhang Z, Qin W, Qian X. A new tandem enrichment strategy for the simultaneous profiling of O-GlcNAcylation and phosphorylation in RNA-binding proteome. The Analyst 2021 146(4) 33465208
Abstract:
RNA-protein interactions play important roles in almost every step of the lifetime of RNAs, such as RNA splicing, transporting, localization, translation and degradation. Post-translational modifications, such as O-GlcNAcylation and phosphorylation, and their "cross-talk" (OPCT) are essential to the activity and function regulation of RNA-binding proteins (RBPs). However, due to the extremely low abundance of O-GlcNAcylation and the lack of RBP-targeted enrichment strategies, large-scale simultaneous profiling of O-GlcNAcylation and phosphorylation on RBPs is still a challenging task. In the present study, we developed a tandem enrichment strategy combining metabolic labeling-based RNA tagging for selective purification of RBPs and HILIC-based enrichment for simultaneous O-GlcNAcylation and phosphorylation profiling. Benefiting from the sequence-independent RNA tagging by ethynyluridine (EU) labeling, 1115 RBPs binding to different types of RNAs were successfully enriched and identified by quantitative mass spectrometry (MS) analysis. Further HILIC enrichment on the tryptic-digested RBPs and MS analysis led to the first large-scale identification of O-GlcNAcylation and phosphorylation in the RNA-binding proteome, with 461 O-GlcNAc peptides corresponding to 300 RBPs and 671 phosphopeptides corresponding to 389 RBPs. Interestingly, ∼25% RBPs modified by two PTMs were found to be related to multiple metabolism pathways. This strategy has the advantage of high compatibility with MS and provides peptide-level evidence for the identification of O-GlcNAcylated RBPs. We expect it will support simultaneous mapping of O-GlcNAcylation and phosphorylation on RBPs and facilitate further elucidation of the crucial roles of OPCT in the function regulation of RBPs.
O-GlcNAc proteins:
NACAM, SAP18, PLOD2, NOP56, DDX3X, PLXB2, RRP8, SERA, PSMD3, MCA3, PRPF3, TPD54, TIM44, ACTN4, ACSL4, PLOD3, IF2P, ZC11A, SC22B, PR40A, MPPB, CSDE1, U520, NU155, EIF3G, SPF27, RL1D1, CLPX, RTN3, LC7L3, VAPB, SMC2, AP2A1, WIZ, BAG2, TOM40, ACL6A, EGFR, LMNA, TFR1, FRIH, RPN1, RPN2, ITB1, SYEP, HNRPC, SRPRA, VIME, GNAI3, ANXA5, LAMP1, ACADM, TOP1, TOP2A, PABP1, ADT3, TPR, EF2, PDIA4, FPPS, ENPL, ALDR, NDKA, RS2, UBF1, ARF4, NUCL, RAB6A, PSB1, FLNA, SDHB, UBA1, NDKB, ITA6, SFPQ, IF4B, AT2B4, THIL, RS12, PSA4, SYVC, 1433T, MAP4, PSA5, PSB4, NDUS1, ECHM, KCY, AMRP, SDHA, METK2, CPSM, PUR9, HNRH1, 1433S, STIP1, P5CR1, MCM4, HSP74, CTNA1, MYH9, DEK, RL4, SPB5, NUP62, RBMX, TCPZ, ECE1, PRS6B, KI67, RAGP1, ATRX, SYQ, LMAN1, NASP, FAS, AL7A1, SYSC, MCM2, ACADV, NU153, RBP2, DNLI3, MRE11, CPT1A, F10A1, TCPD, RAB7A, IDH3G, HCFC1, DHB4, HDGF, ROA3, 6PGD, NUP98, ACLY, TCP4, SYYC, UBP14, SNAA, IF5, TERA, DSRAD, TPD52, EIF3B, NU107, EPIPL, SC61B, SRP54, B2MG, SMD2, RL23A, YBOX1, NOP14, IF4G2, GTF2I, NUCB2, RT22, HMGN5, RBM10, TFAM, CLH1, SPTB2, SET, CAP1, EXOSX, EWS, ODO1, RL18A, NUCB1, M2OM, LMNB2, SRS11, CALD1, RL18, C1QBP, CKAP4, KHDR1, DHX9, GOGA2, SSRP1, AHNK, AIMP1, ILF3, SRSF5, SRSF6, TIF1B, TCOF, PICAL, SNW1, TRI29, EIF3A, MLEC, CAPR1, SMC1A, RRP1B, GANAB, NUMA1, U5S1, RRS1, ACOX1, PLEC, RNPS1, PUM3, RB11B, SEPT7, DDB1, CDC37, SRSF7, PCKGM, HNRL2, INF2, PDS5A, PREP, RRP12, TOIP1, HP1B3, RBM26, BRE1A, CDKAL, PRP8, ZC3HE, QSOX2, IKIP, TM10C, EIF3M, PABP2, KTN1, CAND1, THOC6, P66A, MISP, CCAR1, PELP1, NDUF2, RM50, PAF1, TXND5, TOIP2, THOC2, TM263, NU133, PDC6I, SCFD1, LMO7, ELYS, RT27, HS105, NU205, RAD50, SMRC1, TNPO1, FUBP1, P5CR2, PTCD3, DDX27, EFGM, IWS1, NIBA2, YMEL1, PSMD1, EIF3C, ROAA, CMS1, MBB1A, GNL3, PDIP3, PININ, ACAD9, SFXN1, CYBP, RM47, RTN4, DDX21, AAAS, CARF, AATF, BCLF1, MYOF, SYLC, NXF1, SEC63, LIMA1, SEPT9, KAD3, RCOR1, ACINU, TMCO1, PPIE, PA2G4, RUVB2, TR150, RT23, CHTOP, TLN1, HYOU1, SAM50, SP16H, UTP18, SRPRB
Species: Homo sapiens
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Wang J, Dou B, Zheng L, Cao W, Zeng X, Wen Y, Ma J, Li X. Synthesis of Na(2)S(2)O(4) mediated cleavable affinity tag for labeling of O-GlcNAc modified proteins via azide-alkyne cycloaddition. Bioorganic & medicinal chemistry letters 2021 48 34229054
Abstract:
A facile and convergent procedure for the synthesis of azobenzene-based probe was reported, which could selectively release interested proteins conducted with sodium dithionite. Besides, the cleavage efficiency is closely associated with the structural features, in which an ortho-hydroxyl substituent is necessary for reactivity. In addition, the azobenzene tag applied in the Ac4GlcNAz-labled proteins demonstrated high efficiency and selectivity in comparison with Biotin-PEG4-Alkyne, which provides a useful platform for enrichment of any desired bioorthogonal proteomics.
O-GlcNAc proteins:
PGP, EIFCL, KIF2A, PDLI1, BACH, DFFA, CLIC1, EIF3F, IF2B3, RTCA, PSDE, PPP6, RPC1, PSA7, HNRDL, SC16A, RPAC1, NKRF, EIF3H, PAPS1, SNUT1, ARK72, MYO1B, IDH3B, SAHH2, PLIN3, IMA7, UGDH, CTND1, SNX2, BRD4, WDR1, TBCA, FLNB, PR40A, MPPB, NDUS3, ECI2, CSDE1, U520, WDHD1, EIF3G, PSD10, IDHC, GLRX3, RL1D1, CIAO1, PLPHP, ERLN2, GLSK, SC31A, UBR5, ELP1, VAPB, 6PGL, AGM1, AHSA1, PSMG1, SGPL1, AP2A1, STAU1, TTC4, BPNT1, MBD3, TOM40, ACL6A, GSHR, PNPH, CYTB, KITH, P53, TPM3, PROF1, FUMH, ODPA, CY1, SRP19, DLDH, RU2A, UCHL1, ALDOC, THIO, KAP0, ESTD, ODPB, PYGB, ACADM, G6PD, ADHX, CDK4, HARS1, PEPD, P4HA1, ETFA, MIF, AK1A1, CCNB1, GLNA, DESP, FER, UBF1, PRS6A, RL35A, NELFE, RCC1, E2AK2, SPEE, ANXA7, RAB6A, PSB1, IMDH1, GSTM3, VATB2, FLNA, ACOHC, SDHB, PIMT, FBRL, NDKB, ADRO, TCEA1, TBG1, MAOM, IF4B, THTM, RS12, BRD2, DNJB1, PSA1, PSA2, PSA4, STOM, PYR1, PSB4, PSB6, NDUS1, DPOD1, AMPL, ERP29, PRDX3, ECHM, PEBP1, PDIA3, HMOX2, PURA2, PUR8, AL1B1, RPB2, GDIA, TIA1, QCR1, HNRH3, STIP1, PRDX2, P5CR1, DUT, PROF2, SPB6, RADI, T2FA, MYH9, MYH10, FUS, PRS7, MP2K2, HEM6, GNL1, ODO2, SRP14, TALDO, ETFB, VATA, IF4A3, TXLNA, BUD31, CSK, THIM, LIS1, NAMPT, PRS6B, RECQ1, NOP2, CRKL, NSF, CAPZB, COPD, IDHP, AL9A1, RL34, FAS, SYCC, PSB3, IDH3A, SERPH, ANX11, FXR1, FXR2, SMCA4, GALK1, ROA3, HNRPM, IMA5, GDIR1, HNRPF, KIF11, THOP1, CAZA1, BIEA, MAP11, SUCA, SC24C, DRG2, ECHB, DSRAD, HNRH2, IF6, CORO7, ARPC4, CD81, SC61B, MYL6, PSA6, CDC42, SRP54, UB2D3, UBC12, ARP3, RL37A, COPZ1, NTF2, 1433G, PP1A, PP1B, SMD2, PRS10, ERF1, CNBP, H4, RAP1A, RS30, GBB1, GBB2, TRA2B, 2ABA, DYL1, RL38, PP2AA, TBA1B, GSTO1, DCD, RT05, RT09, RL36A, H33, VIGLN, FKBP3, DHSO, EXOSX, ODO1, MMSA, TF65, LGUL, 1433F, CSTF1, SRS11, EF1A2, PTN11, PUR1, GFPT1, C1QBP, BAX, SRSF4, RBBP4, ASPH, GRSF1, AIMP1, ILF3, CSN1, RED, MTAP, TADBP, ROA0, STX5, SRSF9, SRSF5, IFIT5, EIF3I, DC1I2, PICAL, ULA1, SNW1, FHL1, BOP1, UBP2L, DYHC1, EI2BA, TRI25, FLNC, GNA13, CAPR1, KPRA, UBP10, CHD4, NUMA1, GAPD1, EMC2, SEPT2, IF4H, IPYR, CNN3, SC23B, SF01, TRIP6, MARE1, ELAV1, TOM34, VAMP3, ADRM1, PKN2, CSRP2, DPYL2, RBBP7, H2B2E, PCKGM, TRXR1, TIM50, FA98B, ZN326, PREP, RRP12, SYAM, EXOS6, CAF17, UBR4, NT5D1, PDE12, JMJD6, CDC73, EDC4, PRP8, RL22L, SYDM, GGYF2, HSDL2, TM10C, ZCCHV, DHX29, DCXR, HUWE1, ACOT1, KTN1, CARM1, STX12, HORN, SPB1, SRRM1, SUV3, TXND5, SCPDL, FA98A, PCAT1, FAD1, UBA3, NEK9, BRX1, ZC3HF, SCFD1, HNRLL, ATX2L, PSPC1, P66B, DNJC9, DDX1, H1X, PSMF1, RT27, LAR4B, ARC1A, RENT1, FUBP1, P5CR2, TRM61, ZCCHL, PGAM5, FUBP3, SPF45, THOC3, ZFR, SNX27, RBM14, PRPK, TBCB, CDC5L, PARK7, HCD2, ROAA, EBP2, VRK1, NIPS1, MEP50, TBA1C, ERP44, NTPCR, DDX23, MTNA, NTM1A, TM109, SYTM, THIC, RBM4, HDHD5, ITPA, EIF2A, PDIP3, MK67I, GTPB4, REN3B, API5, UBE2O, WDR12, SLIRP, NAA50, ILKAP, SLK, PININ, YTDC2, RPF2, QTRT2, ARMT1, CSN7B, ELP3, KT3K, MRM3, GLOD4, MCCB, CWC22, WDR6, VTA1, EXOS4, INO1, LUC7L, TIGAR, XPP1, SIAS, PHP14, HELLS, ECHD1, RBM12, DD19A, SEP11, TBC13, ATD3A, DDX18, PNPO, RBM28, LYAR, DPP3, BCLF1, F120A, HPBP1, MAT2B, RRBP1, GMPR2, GRHPR, TES, CHRD1, SEPT9, EI2BD, DBNL, DDX41, APC7, STML2, MRT4, ACINU, NUP50, PSME2, MYO6, CHIP, CSN3, SRRM2, CD11A, SMC3, RTRAF, PIN4, PLAP, NUDC, COF2, AP3M1, TR150, NOP58, SGT1, SYYM, SBDS, EXOS1, SF3B6, RRP15, RT23, STRAP, CHTOP, SAMH1, TLN1, HYOU1, ATG4B, TBL2, PRC2C, PPME1, YTHD2, SNX9, SERC, CLIC4, DC1L1, S23IP
Species: Homo sapiens
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Ramirez DH, Yang B, D'Souza AK, Shen D, Woo CM. Truncation of the TPR domain of OGT alters substrate and glycosite selection. Analytical and bioanalytical chemistry 2021 413(30) 34725712
Abstract:
O-GlcNAc transferase (OGT) is an essential enzyme that installs O-linked N-acetylglucosamine (O-GlcNAc) to thousands of protein substrates. OGT and its isoforms select from these substrates through the tetratricopeptide repeat (TPR) domain, yet the impact of truncations to the TPR domain on substrate and glycosite selection is unresolved. Here, we report the effects of iterative truncations to the TPR domain of OGT on substrate and glycosite selection with the model protein GFP-JunB and the surrounding O-GlcNAc proteome in U2OS cells. Iterative truncation of the TPR domain of OGT maintains glycosyltransferase activity but alters subcellular localization of OGT in cells. The glycoproteome and glycosites modified by four OGT TPR isoforms were examined on the whole proteome and a single target protein, GFP-JunB. We found the greatest changes in O-GlcNAc on proteins associated with mRNA splicing processes and that the first four TPRs of the canonical nucleocytoplasmic OGT had the broadest substrate scope. Subsequent glycosite analysis revealed that alteration to the last four TPRs corresponded to the greatest shift in the resulting O-GlcNAc consensus sequence. This dataset provides a foundation to analyze how perturbations to the TPR domain and expression of OGT isoforms affect the glycosylation of substrates, which will be critical for future efforts in protein engineering of OGT, the biology of OGT isoforms, and diseases associated with the TPR domain of OGT.
O-GlcNAc proteins:
UBA6, SBNO1, CNOT1, SMHD1, ADAS, DX39A, BACH, MYO1C, PSD11, PSD12, TAF4, CLIC1, EIF3F, IPO5, DNM1L, AGRIN, PLOD2, IMA4, NOP56, DDX3X, PDXK, ANM5, TCRG1, PSA7, SCAM3, HAT1, HGS, MYPT1, HNRDL, XPO1, PUR4, NPC1, SCAM2, ARC1B, ARPC2, ARPC3, RPAC1, GNPAT, ZN185, SURF4, OGT1, PPM1G, EIF3D, EIF3H, P4HA2, MAGB2, NUP42, DHX15, MCES, SERA, PSMD3, PAPS1, ZW10, HNRPR, TXNL1, E41L2, XPOT, TIM44, PLRG1, ZN207, GET3, BUB3, ACTN4, AP1G1, SNG1, SNG2, SYNC, MYO1B, NRDC, CALU, EDIL3, SAHH2, DHX16, SMCA5, KDM1A, OGA, HNRPQ, PLOD3, DIAP1, TSN3, PLIN3, CTND1, MAGC1, USO1, DKC1, IF2P, DNJA2, WDR1, CPNE3, T22D2, ANR17, H2AY, FLNB, CISY, SC22B, PR40A, ERLN1, SF3B1, CSDE1, U520, NU155, EIF3G, FACE1, IDHC, ATRN, STAM2, PRAF3, FLOT1, RL1D1, WFS1, MTA2, TOX4, SC24D, SUN1, FRYL, NFAT5, GLSK, SC31A, UBR5, LC7L3, FKBP9, SMC2, IPO7, AHSA1, SGPL1, SC24B, ACSL3, CDS2, AP2A1, BAG2, AIFM1, FADS2, CLPT1, TOM40, ACL6A, LDHA, COX2, PNPH, AATM, PGK1, ASSY, FOS, LMNA, FINC, TFR1, NU4M, NU5M, GBA1, FUCO, ALDOA, ANXA1, OAT, TBB4A, G3P, CPNS1, HSPB1, TYSY, RPN1, RPN2, GNAI2, AT1A1, AT1B1, ALDH2, ADT2, PCCA, IF2A, RLA2, RLA0, LA, ITB1, ATPB, IF4E, ENOA, G6PI, NPM, ITAV, HYEP, LDHB, PDIA1, H10, CATD, ANXA2, CAN1, TBB5, TRY1, SAP, PROF1, SYEP, CATB, HS90A, HNRPC, LAMB1, YES, FUMH, TSP1, ANXA6, MDR1, 4F2, PFKAM, HS90B, SRPRA, ASNS, ODPA, RU17, GNAI3, ANXA5, RSSA, CD63, SNRPA, GSTP1, LEG1, CN37, DLDH, ROA1, PARP1, LKHA4, ALDOC, GPHRB, HS71B, LYAG, RRAS, H14, ODP2, THIO, ESTD, CH60, CLUS, BIP, LAMC1, PPAL, HSP7C, GTR1, GTR3, ODPB, PYGB, LAMP1, TOP1, TOP2A, G6PD, PYC, C1TC, MPRI, ADHX, PABP1, PCNA, HARS1, IMDH2, TPR, KCRB, ACTN1, PEPD, XRCC6, XRCC5, LAMP2, RINI, EF2, PDIA4, P4HA1, TCTP, ACPH, ETFA, GLU2B, FPPS, KPYM, ENPL, IDE, HNRPL, SYDC, ALDR, B4GT1, EZRI, FOSL2, NDKA, GNS, RS2, RFA2, BGAL, H12, AT2A2, FAAA, AATC, JUNB, ITA2, CAN2, PYRG1, DDX5, PFKAL, PRS6A, TCPA, RL35A, ARF4, RL7, VINC, SON, RL17, PGAM1, DNLI1, PLCG1, NUCL, HXK1, RPB3, E2AK2, TFE3, SPEE, IF2B, ANXA7, PSB1, MPRD, LMNB1, VATB2, FLNA, VDAC1, TGM2, FBRL, PUR2, PUR6, UBA1, NDKB, ADRO, ROA2, RFX1, QCR2, SFPQ, PPIB, SYWC, RS3, SAHH, COF1, IF4B, PPAC, RPB1, MCM3, RS12, DNJB1, ATPA, PSA1, PSA2, PSA3, PSA4, ITA3, MOES, DDX6, DNMT1, U2AF2, RL13, PTBP1, SYTC, SYVC, EF1G, STOM, RL10, RFA1, APEX1, CD82, PYR1, CALR, MAP4, CALX, PSB8, PSA5, PSB4, PSB6, PSB5, DPOD1, TEAD1, MK01, AMPL, IMPA1, EPHA2, TKT, EF1D, PRDX6, BLVRB, PRDX5, PRDX3, RL12, ECHM, PEBP1, PDIA3, 2AAA, CDC27, PURA2, AMRP, PUR8, CLIP1, CTR1, RPB2, SDHA, GDIA, METK2, DNJA1, QCR1, PUR9, HNRH3, HNRH1, 1433B, 1433S, STIP1, PRDX2, RL9, KINH, MRP1, MCM4, MCM5, MCM7, GLYM, HSP74, CTNA1, CTNB1, PHB1, RADI, RFC4, RL22, FBN2, GDE, MYH9, MYH10, COPB2, SOAT1, ADDA, BASI, FUS, NU214, PRS7, MP2K2, ATPG, HEM6, RL4, PGM1, ODO2, SRP14, NUP62, FDFT, TAGL2, TALDO, RBMX, VKGC, VATA, GRP75, IF4A3, RS19, RL3, OST48, AN32A, CAP2, TCPZ, RL13A, STAT3, MDHC, MDHM, ECHA, IF2G, KDM5C, GARS, SYIC, STAT1, CASP3, LPPRC, SATT, LIS1, MUC18, MATR3, MSH2, MAGA4, RANG, NAMPT, PRS6B, PPIC, VDAC2, UBP5, KI67, RAGP1, RECQ1, NOP2, CRKL, BAG6, NSF, RL27A, RL5, RL21, RL28, RS9, RS5, MAP1B, UTRN, IQGA1, STT3A, CAPZB, SYQ, UCRI, PPCE, COPD, LSS, CD151, PSMD8, TCPE, NEST, HSP13, PIPNB, PAXI, AL9A1, RL34, LMAN1, NASP, FAS, TCPG, EFTU, AL7A1, SYAC, SYCC, SYSC, PRI2, PSB3, PSB2, MCM2, ACADV, YLPM1, TMEDA, RBM25, NU153, RBP2, NDUV1, GSK3A, TAF6, GUAA, GDIB, EMD, SERPH, VASP, TCPQ, TCPD, ANX11, PAPOA, FXR1, RAB5C, RAB7A, RT29, SMCA4, BAP31, HCFC1, DHB4, HDGF, ROA3, 6PGD, HNRPM, IMA1, GDIR1, AGFG1, HNRPF, MSH6, RBM5, SPSY, CAZA1, NUP98, BIEA, PPP5, ACLY, MAP11, COPB, COPA, SC5A3, SMTN, MOT1, IST1, SC24C, SYRC, SYYC, UBP14, BCAT1, AT1B3, RD23B, P5CS, IF5, S12A2, XPO2, TERA, ECHB, AFAD, NP1L1, DSRAD, SEC13, PSA, HNRH2, EIF3B, ATPK, SYMC, IF6, CTBP2, TMM33, GEMI4, CORO7, NU107, ARPC4, TPIS, EIF3E, PP4C, IF4A1, PSA6, CDC42, DEST, GMFB, SRP54, RAB10, UBC12, ARP3, ARP2, ABCE1, RAP2B, RS3A, RL15, RL37A, S61A1, DAD1, NPC2, HNRPK, 1433G, RS7, PRS4, PRS8, RS8, RS15A, RS16, 1433E, RS14, RS23, RS13, RS11, SMD2, SMD3, PRS10, RL7A, ERF1, CNBP, RS4X, PP2AB, RS6, H4, RAB1A, RAN, RL23, RS25, RS28, RL30, RL31, RL10A, RL32, RL11, RL8, PPIA, TRA2B, AP2B1, 1433Z, IF5A1, RACK1, YBOX1, UB2L3, TBA4A, IF4G2, GTF2I, TCPB, PRKDC, RL24, SRSF3, FOXK1, DAB2, RBM10, S25A3, VIGLN, CLH1, FKBP3, HNRPU, U2AF1, SPTB2, AMPD2, CAP1, HMCS1, LAT1, EXOSX, PFKAP, EWS, PLCB3, ODO1, RL18A, MP2K1, FKBP4, PLOD1, RL6, TOP2B, DYST, KMT2A, LMNB2, CEBPZ, TF65, IF4G1, LGUL, 1433F, DYN1, EF1A2, PTN11, PUR1, GFPT1, EXOS9, SUH, GABPA, PRDX1, RL18, C1QBP, CKAP4, KHDR1, LRP1, SRSF1, PP2BA, DHX9, GOGA2, LG3BP, CD47, SSRP1, NSUN2, NCBP1, EP300, AHNK, MGAT2, GALT2, AP1B1, MPPA, SCRN1, NU160, SCAP, TBL3, TF3C1, ASPH, TROP, BPTF, AIMP1, ILF2, ILF3, LMAN2, TRAP1, FLII, ACACA, CSN1, MTAP, EI2BE, TADBP, ROA0, PRDX4, PSMD2, SRSF6, TIF1B, DHRS2, G3BP1, MTA1, EIF3I, UB2V1, DC1I2, NNTM, TCOF, TMED1, VPP3, PICAL, RIPK1, CUL2, CUL4A, CUL4B, FCL, MOGS, CD166, SPTN1, DX39B, BLMH, CKAP5, COTL1, HNRPD, SCRB2, EIF3A, UBP2L, SCRIB, FHL2, DPYL3, DCTN1, DYHC1, SRC8, FLOT2, TRI25, FLNC, GALE, CAPR1, RBM39, MCM6, TRIPC, PUM1, EPN4, RRP5, BMS1, UBP10, GANAB, LBR, MVP, MEF2D, CHD4, LASP1, GPNMB, ZN638, IMB1, NUMA1, CND2, SPCS2, PSMD6, ABRX2, SEPT2, CND1, U5S1, SYK, WDR43, PDIA6, PA1B3, IPYR, TEBP, NONO, PWP2, RCN1, L2GL1, TMED2, PCBP1, PCBP2, SF3B3, SC23A, SF3A1, TSN, SF01, PCH2, MED1, MARE1, ELAV1, NSDHL, TAB1, AAAT, MLF2, VAS1, ZYX, SEPT7, ADRM1, CCDC6, UAP1, PSMD5, PKN2, DDB1, DPYL2, KCC4, SRSF7, DREB, NRF1, FSCN1, IF16, PCKGM, UPP1, UGPA, TRXR1, HNRL2, INF2, PDS5A, QSER1, QRIC1, SMU1, P3H1, K2C71, TIM50, GXLT1, TB10B, SNUT2, HERC4, P4R3B, EOGT, CE170, UBR4, SKT, UBAP2, RBM26, RPRD2, ZMYM4, TWF2, LIN54, RHBD2, MBOA5, EDC4, PRP8, SCYL2, NFRKB, SKI3, NCEH1, BRAT1, ZC3HE, C1TM, SBSN, CREL2, PACS1, GGYF2, SRCAP, TMTC3, IKIP, SPT6H, SND1, DDX46, 5MP2, DHX30, EIF3M, CYFP1, KDM3B, RIR2B, ERMP1, ZCCHV, NUP54, POGZ, MYH14, CLAP1, EMSY, SH3R1, HUWE1, TMED4, SYVN1, ZN598, PHLB1, LUZP1, CAND1, TSYL5, EZHIP, CARM1, DDX42, P66A, C2CD5, I2BP1, RB6I2, PLD3, P3H3, WDR75, Z3H7A, ANKH1, CCAR1, DJC10, SMAP1, SPART, RPTOR, NUP93, ZN687, SYNPO, FNBP4, CPSF7, EMC1, ENAH, S43A3, GT251, SUMF2, PGLT1, TXND5, SERB1, LS14A, ABCF1, FBX22, NEUA, CADM4, ATLA2, PLBL2, WDR36, SMRC2, NPL4, S35B2, UBA3, TBC15, CIP2A, STT3B, PNPT1, HM13, DTX3L, PO210, GEMI5, PARD3, IPO4, SCRB1, RAB2B, NU133, PDC6I, S20A1, ATX2L, PALLD, DDX1, GBF1, NICA, TM9S4, HS105, GCN1, PRP16, NU205, PXDN, TFG, ARC1A, HDAC2, CBP, SYMPK, DDX17, CELF1, UFD1, GSLG1, RENT1, TNPO1, GLMN, USP9X, UBP7, SCAM4, SMCE1, MYDGF, PIGT, NCLN, EXOC4, FERM2, FUBP1, FKB10, APH1A, PPWD1, AP2M1, RCN3, ELP4, SEH1, DNJA3, CYFP2, PGM2, PDLI5, ERO1A, FUBP3, ITCH, TMX3, CNDP2, ZFR, EP400, SNX27, PRRC1, ZN512, MBOA7, DOCK7, ARAP1, IPO9, RBM14, NED4L, VPS35, CIC, MED15, EFGM, MCCA, MYADM, SIN3A, MINT, PNKP, NIBA2, TBCB, PSB7, CNN2, PSMD1, EYA3, SORT, NUP88, P2RX4, DNJC7, PHB2, KIF2C, ATX2, HCD2, ROAA, NP1L4, ACON, TM9S2, S29A1, TS101, TCPH, ANM1, CND3, MBB1A, COR1B, GCP2, ESYT1, DIDO1, MCMBP, TBCD, TBB6, RPC3, HNRL1, HTAI2, TMED9, NUP58, SFXN3, OSB10, NAA15, S12A9, SRRT, EIF2A, WDR11, FTO, TTYH3, WDR12, YIPF3, PITH1, PAIP1, NAT10, XRN2, TOLIP, SP130, BRD8, DDX47, MAGT1, RAB1B, LMA2L, I2BPL, GHITM, TMX1, PTN23, OFUT1, PIGU, WNK1, AMPB, HEAT1, NOL6, WDR26, AT133, ACAD9, GORS2, MARH7, SFXN1, PKHA5, XPO5, MYG1, EMAL4, TM9S3, APMAP, ARFG3, RTN4, ANLN, XPP1, PDLI7, DDX21, EI2BG, MBNL1, HEBP1, SERC1, F1142, SYFB, SYIM, SMC4, OLA1, RBM12, T106B, CYRIB, DCA13, CC50A, SEP11, FANCI, ARL8B, DDX18, TM160, MIO, CARF, DPP3, RM39, UGGG1, ERAP1, HPBP1, MAT2B, MYOF, ITSN2, HACD3, SEP10, RCC2, DIP2B, CT2NL, RRBP1, SYLC, RBM27, SUCB1, MRC2, DHCR7, DJB11, SAE2, NXF1, SUN2, SRP68, SEPT9, UBQL2, PCYOX, CEIP2, S30BP, MED13, PUF60, NRBP, VATH, IPO11, XPO7, BAZ2A, GGT7, NAGK, DBNL, CDC23, STML2, PARP4, NUDT5, TF3C4, ACINU, RAB21, DNPEP, ASAP1, MRTFB, MED23, HECD1, ZMYD8, COR1C, MYO6, PRP19, MAGD2, PSD13, MINP1, NOVA2, WDR3, S12A4, MACF1, SCAF8, TRI33, SMAG1, XCT, SRRM2, PA2G4, SMC3, RUVB2, EIF3L, PLAP, RUVB1, VDAC3, ERGI3, SC5A6, DRG1, NCKP1, AT11B, E41L3, CHSP1, NOP58, ZN281, SYYM, TMX2, SHLB1, SBDS, TMED7, STRAP, RTCB, FBX7, RBGP1, NOC2L, TLN1, DCAF1, HYOU1, PRC2C, YTHD2, SP16H, TNPO3, MAGD1, SNX8, PUS1, NPTN, S12A7, COPG1, BIG1, 5MP1, DC1L1, IF2B2, ZNT1, S4A7, ZHX2, S23IP
Species: Homo sapiens
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Ramirez DH, Aonbangkhen C, Wu HY, Naftaly JA, Tang S, O'Meara TR, Woo CM. Engineering a Proximity-Directed O-GlcNAc Transferase for Selective Protein O-GlcNAcylation in Cells. ACS chemical biology 2020 15(4) 32119511
Abstract:
O-Linked β-N-acetylglucosamine (O-GlcNAc) is a monosaccharide that plays an essential role in cellular signaling throughout the nucleocytoplasmic proteome of eukaryotic cells. Strategies for selectively increasing O-GlcNAc levels on a target protein in cells would accelerate studies of this essential modification. Here, we report a generalizable strategy for introducing O-GlcNAc into selected target proteins in cells using a nanobody as a proximity-directing agent fused to O-GlcNAc transferase (OGT). Fusion of a nanobody that recognizes GFP (nGFP) or a nanobody that recognizes the four-amino acid sequence EPEA (nEPEA) to OGT yielded nanobody-OGT constructs that selectively delivered O-GlcNAc to a series of tagged target proteins (e.g., JunB, cJun, and Nup62). Truncation of the tetratricopeptide repeat domain as in OGT(4) increased selectivity for the target protein through the nanobody by reducing global elevation of O-GlcNAc levels in the cell. Quantitative chemical proteomics confirmed the increase in O-GlcNAc to the target protein by nanobody-OGT(4). Glycoproteomics revealed that nanobody-OGT(4) or full-length OGT produced a similar glycosite profile on the target protein JunB and Nup62. Finally, we demonstrate the ability to selectively target endogenous α-synuclein for O-GlcNAcylation in HEK293T cells. These first proximity-directed OGT constructs provide a flexible strategy for targeting additional proteins and a template for further engineering of OGT and the O-GlcNAc proteome in the future. The use of a nanobody to redirect OGT substrate selection for glycosylation of desired proteins in cells may further constitute a generalizable strategy for controlling a broader array of post-translational modifications in cells.
O-GlcNAc proteins:
SBNO1, CNOT1, P121C, DX39A, GTPB1, AP3B1, PGRC1, TAF4, EIF3F, IPO5, IF2B3, NOP56, DDX3X, ARI1A, IRS4, ANM5, TCRG1, PSA7, HGS, MYPT1, HNRDL, XPO1, SET1A, PUR4, NPC1, TIF1A, NKRF, OGT1, PPM1G, EIF3D, EIF3H, DHX15, SERA, HNRPR, IF4G3, E41L2, ZN207, BUB3, ACTN4, HTSF1, AP1G1, SYNC, AKAP8, CALU, SMCA5, JIP4, OGA, HNRPQ, DIAP1, TSN3, SNX2, DKC1, CLAP2, CPNE3, PHF2, ANR17, H2AY, FLNB, NCOR1, CISY, PR40A, SF3B1, CSDE1, U520, EIF3G, PRAF3, SRP72, MTA2, TOX4, SC24D, SC31A, SCAF4, ZRAB2, LC7L3, VAPB, IPO7, SC24B, ACSL3, AP2A1, AIFM1, LDHA, COX2, HPRT, AATM, PGK1, LMNA, TFR1, ALDOA, OAT, G3P, RPN1, RPN2, AT1A1, ADT2, PCCA, IF2A, RLA0, ITB1, ATPB, ENOA, PYGL, G6PI, NPM, LDHB, PDIA1, H10, TBB5, HEXB, PROF1, SYEP, HS90A, HNRPC, 4F2, HS90B, ASNS, ODPA, RU17, RSSA, SNRPA, GSTP1, HMGB1, DLDH, ROA1, PARP1, LKHA4, HS71B, H14, ODP2, THIO, CH60, BIP, HSP7C, EPB41, ODPB, LAMP1, ACADM, TOP1, TOP2A, PYC, C1TC, MPRI, PRPS2, PABP1, PCNA, HARS1, IMDH2, TPR, KCRB, XRCC6, XRCC5, EF2, PDIA4, PLST, GLU2B, KPYM, ENPL, PO2F1, HNRPL, SYDC, PLAK, ALDR, EZRI, GNS, RS2, CREB1, H12, AT2A2, JUNB, PYRG1, DDX5, PRS6A, TCPA, RL35A, RL7, VINC, SON, RCC1, NUCL, HXK1, E2AK2, SPEE, IF2B, ANXA7, LMNB1, FLNA, VDAC1, FBRL, PUR2, PUR6, UBA1, NDKB, ROA2, RFX1, TCEA1, SFPQ, PPIB, RS3, NFYA, SAHH, COF1, IF4B, EF1B, MCM3, BRD2, ATPA, PSA1, PSA3, PSA4, PAX6, U2AF2, RL13, PTBP1, SYTC, SYVC, EF1G, RFA1, APEX1, PYR1, CALR, MAP4, CALX, PSB5, TKT, PRDX6, PRDX5, PRDX3, RL12, PEBP1, PDIA3, 2AAA, CDC27, AMRP, SDHA, QCR1, PUR9, HNRH1, STIP1, PRDX2, RL9, CSTF2, MCM4, MCM5, MCM7, GLYM, HSP74, PHB1, MYH9, COPB2, ADDA, BASI, FUS, NU214, DEK, MP2K2, ATPG, RL4, SRP14, NUP62, RBMX, GRP75, IF4A3, RS19, RL3, TXLNA, TCPZ, MDHC, MDHM, ECHA, IF2G, GARS, SYIC, LAP2A, LAP2B, MTREX, RS27, LPPRC, MATR3, RANG, VDAC2, UBP5, KI67, RAGP1, NOP2, CRKL, BAG6, RL27A, RL5, RL21, RL28, RS9, STT3A, COPD, PRC2A, TCPE, AL9A1, RL34, NASP, FAS, TCPG, EFTU, SYAC, SYSC, PSB3, MCM2, YLPM1, TMEDA, RBM25, NU153, RBP2, GSK3A, TAF6, GUAA, MRE11, GDIB, EMD, F10A1, LRBA, RL14, TCPQ, TCPD, ANX11, PAPOA, RAB7A, SMCA4, HCFC1, DHB4, ROA3, 6PGD, HNRPM, IMA1, AGFG1, HNRPF, MSH6, RBM5, NUP98, ACLY, COPB, COPA, MOT1, SC24C, SYRC, SYYC, AT1B3, RD23B, P5CS, IF5, XPO2, TERA, AFAD, DSRAD, PSA, SYMC, CTBP2, NU107, TPIS, ACTB, IF4A1, PSA6, ARF3, ABCE1, RAP1B, RS3A, RL26, RL15, S61A1, HNRPK, RS7, RS8, 1433E, RS14, RS23, RS11, SMD1, RL7A, RS4X, H4, RAN, RL23, GBB2, RL10A, RL11, RL8, PPIA, RL40, TRA2B, AP2B1, IF5A1, RACK1, YBOX1, EF1A1, TBB4B, CSK21, IF4G2, GTF2I, TCPB, PRKDC, RL24, RL19, SRSF3, FOXK1, RBM10, S25A3, CLH1, HNRPU, SPTB2, FOXK2, CAP1, LAT1, EXOSX, EWS, RL18A, FKBP4, RL6, TOP2B, KMT2A, LMNB2, TF65, IF4G1, TLE3, SRS11, PUR1, SUH, GABPA, PRDX1, RL18, C1QBP, KHDR1, SRSF1, DHX9, CD47, SSRP1, RBBP4, AHNK, AP1B1, NU160, BPTF, TP53B, AIMP1, ILF2, ILF3, LMAN2, TRAP1, PP1R8, ACACA, ROA0, PRDX4, CBX3, PSMD2, SRSF6, TIF1B, PTK7, PABP4, EIF3I, TCOF, SF3B2, TMED1, PICAL, RIPK1, HDAC1, CUL4B, CD166, IDI1, NFYC, CKAP5, HNRPD, SCRB2, DSG2, EIF3A, UBP2L, SCRIB, TTL12, DCTN1, DYHC1, SRC8, CAPR1, RBM39, MCM6, MDC1, EPN4, SMC1A, RRP1B, UBP10, GANAB, LBR, ZN638, IMB1, NUMA1, SEPT2, SART3, U5S1, SYK, BRD3, PDIA6, IPYR, TEBP, NONO, PWP2, RCN1, PCBP1, PCBP2, SF3B3, SAFB1, SF3A1, NCOA2, SF01, MARE1, NSDHL, TAB1, AAAT, VAS1, ZYX, CCDC6, PKN2, DDB1, CDC37, SRSF7, CPSF6, NRF1, H31T, QSER1, QRIC1, P3H1, TB10B, AMOT, DHB12, PRC2B, H2B2F, HP1B3, CE170, ZC3HD, RBM26, RIF1, RPRD2, ZN318, ECM29, ZMYM4, MAP1S, LIN54, EDC4, PRP8, SCYL2, NFRKB, ZC3HE, LARP1, FIP1, MCAF1, GGYF2, SPT6H, SND1, DHX30, KDM3B, ZCCHV, NUP54, POGZ, NUFP2, MAVS, I2BP2, RBBP6, HUWE1, YTHD3, CENPV, LYRIC, ZN598, GP180, CAND1, CARM1, DDX42, P66A, ARI3B, MGAP, PHF6, CHERP, ANKH1, SUGP1, CCAR1, SPB1, PHAR4, SPART, CCAR2, NUP93, S11IP, FNBP4, CPSF7, ARFG1, ENAH, SPAT5, TXND5, LS14A, Z280C, TNR6A, SMRC2, TBC15, PNPT1, HM13, PO210, GEMI5, ZN384, SMAP2, NU133, PDC6I, PCNP, CKAP2, ATX2L, P66B, ELYS, DDX1, GBF1, NICA, UBXN4, HS105, LAR4B, NU205, AKAP1, TFG, CBP, DDX17, CELF1, RENT1, SMRC1, FUBP2, TNPO1, UBP7, NCLN, FUBP1, FKB10, KBP, PDLI5, FUBP3, CHAP1, Z512B, ZFR, PRRC1, DOCK7, RBM14, VPS35, CIC, EFGM, SIN3A, MINT, CDC5L, PSMD1, EYA3, ATX2, HCD2, ACON, TS101, TCPH, ANM1, SH3G1, COR1B, DIDO1, HNRL1, DDX23, TMED9, NUP58, RBM4, NAA15, B2L13, YTHD1, UNK, ILKAP, SP130, BRD8, I2BPL, SLK, S6A15, PININ, NELFA, PTN23, WNK1, AMPB, GORS2, CYBP, TAF9B, GLOD4, CBX8, NCOA5, CHD8, APMAP, DCP1A, RTN4, ANLN, GEPH, PDLI7, DDX21, SYFB, SYIM, SMC4, RBM12, DDX18, CARF, UGGG1, CDK12, TECR, IF2B1, HPBP1, ITSN2, CNOT2, HACD3, RCC2, SYLC, SUCB1, UBQL2, PCYOX, S30BP, PUF60, NRBP, DACH1, BAZ2A, BAZ1B, CDC23, TASOR, ACINU, CDV3, MRTFB, YETS2, HECD1, ZMYD8, DD19B, PRP19, MAGD2, FAF1, TRI33, SRRM2, PA2G4, RUVB2, RUVB1, VDAC3, E41L3, TR150, NOP58, SHLB1, LC7L2, TMED7, STRAP, RTCB, HBS1L, TLN1, HYOU1, PRC2C, SP16H, COPG1, DC1L1, S23IP
Species: Homo sapiens
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Zhang W, Liu T, Dong H, Bai H, Tian F, Shi Z, Chen M, Wang J, Qin W, Qian X. Synthesis of a Highly Azide-Reactive and Thermosensitive Biofunctional Reagent for Efficient Enrichment and Large-Scale Identification of O-GlcNAc Proteins by Mass Spectrometry. Analytical chemistry 2017 89(11) 28510447
Abstract:
O-linked β-N-acetylglucosamine (O-GlcNAc) is a ubiquitous post-translational modification of proteins in eukaryotic cells. Despite their low abundance, O-GlcNAc-modified proteins play many important roles in regulating gene expression, signal transduction, and cell cycle. Aberrant O-GlcNAc proteins are correlated with many major human diseases, such as Alzheimer's disease, diabetes, and cancer. Because of the extremely low stoichiometry of O-GlcNAc proteins, enrichment is required before mass spectrometry analysis for large-scale identification and in-depth understanding of their cellular function. In this work, we designed and synthesized a novel thermosensitive immobilized triarylphosphine reagent as a convenient tool for efficient enrichment of azide-labeled O-GlcNAc proteins from complex biological samples. Immobilization of triarylphosphine on highly water-soluble thermosensitive polymer largely increases its solubility and reactivity in aqueous solution. As a result, facilitated coupling is achieved between triarylphosphine and azide-labeled O-GlcNAc proteins via Staudinger ligation, due to the increased triarylphosphine concentration, reduced interfacial mass transfer resistance, and steric hindrance in homogeneous reaction. Furthermore, solubility of the polymer from complete dissolution to full precipitation can be easily controlled by simply adjusting the environmental temperature. Therefore, facile sample recovery can be achieved by increasing the temperature to precipitate the polymer-O-GlcNAc protein conjugates from solution. This novel immobilized triarylphosphine reagent enables efficient enrichment and sensitive detection of more than 1700 potential O-GlcNAc proteins from HeLa cell using mass spectrometry, demonstrating its potential as a general strategy for low-abundance target enrichment.
O-GlcNAc proteins:
A0A024R5Q9, A0A0B4J1V8, A0A0B4J203, GAL3B, RBM47, UBA6, ESYT2, SHOT1, NDUF8, VWA8, GTPBA, A5Y5A3, CNOT1, PGP, TBAL3, SMHD1, LR14B, NCF1B, A8K005, B4DLC0, B4DLN1, B4DQP1, B4E259, EIFCL, B7Z2B5, B7Z532, B7Z5W1, DSA2D, D7RF68, E9PL57, E9PLN8, I1VE18, L0R5A1, S35U4, ADAS, KIF2A, PDLI1, AIP, SMAP, NDUS8, PSD12, PSMD9, SPT5H, DFFA, HIP1, EIF3F, IPO5, SAP18, IF2B3, DNM1L, NDUA4, PSDE, BIN1, NOP56, DDX3X, IMA3, TRI38, SCD, DHSD, TXND9, ACPM, UBFD1, AP3D1, DVL2, CHD1, TOR1A, PTGES, ACOT8, NHRF1, TPP1, RPC1, PSA7, MGST3, HAT1, QCR8, HGS, S27A2, HNRDL, BTAF1, SR140, PUR4, NPC1, DEGS1, SCAM2, ARPC2, ARPC3, BET1, RPAC1, PGRC2, NDUA1, RER1, OGT1, PMM2, HMGB3, EIF3H, ARPC5, DHX15, CYB5B, PRP4, SERA, NDUS4, PAPS1, ZW10, SNUT1, RIPK2, PRPF3, PPIH, E41L2, XPOT, DNPH1, PSCA, TSN6, ZN207, NDUB5, BUB1, BUD23, GATC, HTSF1, SGTA, SYNC, PM34, CPSF5, SCO2, ORC5, TIM8A, DHX16, SMCA5, MAST3, OPA1, TBCD4, OGA, HNRPQ, DIAP1, MOT2, IMA7, CTND1, DPM1, Z354A, RT14, PQBP1, TI17B, DKC1, IF2P, EDF1, MCE1, ABCB7, WDR1, BRE1B, ZC11A, CLU, ANR17, COQ9, NDUS7, NIPS2, VPS4B, CISY, PR40A, NDK6, VP26A, MPPB, PSIP1, ECI2, BAF, SF3B1, CSDE1, PRKRA, NPM3, ERAL1, TIPRL, KS6A4, NU155, XRP2, WDHD1, CRTAP, S2512, RNH2A, EIF3G, AP1G2, IDHC, CCNK, SPF27, FLOT1, GLRX3, RL1D1, CLPX, CIAO1, SRP72, LTN1, UFL1, UBXN7, PRP6, PCF11, GLSK, SC31A, RPP14, YIF1A, AGFG2, NDUB6, UBE4B, ELP1, NDUB8, LC7L3, KIF4A, MPZL1, 6PGL, PAPS2, TRI16, ARI2, PSMG1, MPC2, ASML, RPP29, HS74L, AP2A1, TTC4, TM50A, BAG2, EMAL2, BPNT1, NDUBA, ACL6A, COX1, COX2, PNPH, AATM, EGFR, ADA, ATP6, RASN, LDLR, HLAB, LMNA, ALBU, NU1M, NU3M, NU4M, KITH, K2C1, G3P, HLAA, TYSY, RPN2, HMGN1, ISG15, LA, K1C18, DCUP, CDK1, NPM, TPM3, HEXA, HYEP, H10, SAP, SYEP, HNRPC, YES, SP1, DAF, PFKAM, HEM3, CY1, RU17, ITA5, RS17, RSSA, RM03, SRP19, RU1C, HMOX1, RU2A, PARP1, LKHA4, ALDOC, F86B2, GSTT2, ST1A3, HS71B, RPAC2, ECE2, EFMT4, EFCE2, MYBA, HLAC, ARAF, H14, ODP2, MGST1, ESTD, GTR3, UMPS, ODB2, ACADM, TOP1, TOP2A, G6PD, PYC, C1TC, ADHX, PRPS2, PABP1, ADT1, TPR, KCRB, RINI, K2C5, ACPH, GYS1, ENOB, CD59, MTDC, MIF, CX7A2, CCNB1, CX6B1, PO2F1, SYDC, PLAK, QCR7, ERF3A, EZRI, UCHL3, MCP, PHKG2, RS2, RFA2, CBR1, BGAL, EPCAM, NCPR, AT2A2, UBF1, NDUB7, KAPCA, CAN2, PYRG1, DDX5, LEG3, TCPA, PTN1, RL7, NELFE, PGAM1, RCC1, XRCC1, RPAB1, NDUV2, SPEE, IF2B, BTF3, H2A1D, COX5A, LMNB1, IMDH1, FLNA, ACOHC, ODBB, MUTA, OSBP1, PIMT, FBRL, PUR2, HEMH, TCEA1, AT2B4, CPT2, KTHY, RIR1, EF1B, AT5F1, THIL, CDK2, APC, MCM3, THTM, RS12, TYY1, ITA3, DDX6, DNMT1, U2AF2, HMGB2, SYVC, EF1G, AOFB, RL10, RFA1, APEX1, MAP4, CALX, PSA5, NDUS1, RXRB, AMPL, T2EA, T2EB, IMPA1, 3MG, CO4A5, SPB3, ATPD, PDIA3, 2AAB, CDC27, NMT1, PURA2, AMRP, PUR8, CLIP1, GSTT1, CTR1, AL1B1, RPB2, METK2, RIR2, HNRH3, HNRH1, STIP1, L1CAM, P5CR1, TF2H1, ACSL1, CSTF2, MCM4, MCM5, MCM7, RFC4, RFC2, RL22, T2FA, GDE, SOAT1, BASI, NU214, DEK, K22E, PRS7, HEM6, RL4, LONM, SRP14, NUP62, TALDO, RBMX, COIL, RS19, FEN1, CAP2, TCPZ, NNMT, PSB10, RL13A, PEX19, MDHM, RFC5, RFC3, BUD31, GARS, SYIC, EIF1, LAP2A, LAP2B, MTREX, AK1C3, LPPRC, THIM, MATR3, GPDM, SSRA, SSRB, RANG, PRS6B, EFTS, VDAC2, UBP5, RECQ1, ATRX, IF2M, MP2K3, BRCC3, RL27A, RS10, GNPI1, CAPZB, IF1AX, SYQ, RL29, MAOX, PTSS1, IDHP, PIPNB, DPOD2, TCPG, RM19, INPP, SRP09, SYCC, PSB2, MCM2, ACADV, TMEDA, RBM25, NU153, RBP2, NDUV1, GSK3B, 5NTC, SPS1, GUAA, DNLI3, GDIB, CPT1A, F10A1, DYN2, MAP2, CDK7, CDK9, LRBA, RL14, PAPOA, FXR1, FXR2, RAB7A, RB27A, RT29, SMCA4, SSRD, TPMT, HCFC1, SSDH, DHB4, PSMD7, STA5B, UBP11, DYLT3, CCNH, ROA3, HNRPM, IMA1, NCBP2, MP2K6, KIF11, HXK2, BIEA, PLK1, NUBP1, MAP11, SUCA, MVD1, PGTB2, CATC, CCHL, IST1, TCP4, PMS2, SYYC, UBP14, RD23A, P5CS, IF5, PSMD4, BI1, MFAP1, AFAD, NP1L1, CASP9, ADK, DSRAD, SEC13, HNRH2, SCOT1, EIF3B, ATP5E, IF6, CF298, TMM33, CORO7, SELB, MTPN, YBEY, ARPC4, CD81, SC61B, ROMO1, MYL6, ACTB, IF4A1, PRPS1, PSA6, CDC42, DEST, GMFB, RAB8A, SPCS3, SRP54, RAB5B, UBC12, UBE2K, RAB14, TM258, CSN2, RAP1B, RS3A, VA0D1, NAA20, S61A1, B2MG, COPZ1, UFM1, AP1S1, HNRPK, TIM10, PRS4, RS15A, RS23, RUXE, SMD3, PRS10, ERF1, RS6, RAN, RL23, RS15, RS24, RS25, RS26, RBX1, RL39, RL31, RL10A, RL32, PPIA, TRA2B, AP2B1, DYL1, DYLT1, RL38, SKP1, ACTG, UBC9, CSK2B, UB2L3, EF1A1, CSK21, SERB, IF4G2, RPP38, GTF2I, RAE1L, GSTO1, PRKDC, IN35, RT25, RT35, RT05, RT11, RT15, RT06, RT09, LACTB, RL36A, ARF5, ERH, H33, FOXK1, PRR5, RBM10, TFAM, PIPNA, TF2B, CDK16, NFKB2, FKBP3, HNRPU, U2AF1, SET, GALK2, FABP5, CAP1, HMCS1, IFM3, RL18A, NUCB1, RL6, M2OM, LMNB2, TAP1, TAP2, CEBPZ, TF65, UBXN1, IF4G1, ATP7A, CSTF1, FAK1, SRS11, EF1A2, PUR1, PSME1, GABPA, FMR1, RL18, C1QBP, KHDR1, KLC1, SRSF1, QOR, LG3BP, PPID, SSRP1, CAMP2, NSUN2, RBBP4, NCBP1, HSP7E, BST2, MPPA, WASC5, TBL3, SNF5, STX4, GRSF1, NFIA, SF3A3, TP53B, ILF2, ILF3, PTPRJ, TRAP1, TRAF2, MYO1E, PP1R8, BNIP2, CSTF3, STRN3, RM28, ACACA, CSN1, CAF1B, TADBP, ROA0, AIMP2, PAK2, CBX3, STK3, STX5, SRSF5, SRSF6, TIF1B, PABP4, EIF3I, 2A5G, UB2V1, RM49, DC1I2, ILK, SNTB2, PPIG, TCOF, SF3B2, HAP28, ROCK1, PICAL, SQSTM, PRP4K, HDAC1, DCTN2, SNW1, STIM1, PWP1, MTMR2, CUL2, CUL4A, FCL, RAB31, RAB32, NOG2, TBB2A, BYST, PEBB, HNRPD, IL18, VEZF1, TRI29, TTL12, DPOA2, ICT1, DCTN1, ELOA1, TRI25, FASTK, PDE3A, RBM39, HLTF, MCM6, ITPR1, TRIPC, PUM1, EPN4, SMC1A, RRP1B, PSF1, UBP10, GANAB, LBR, CHD4, NOLC1, NUMA1, PSME4, GAPD1, CND2, SPCS2, EMC2, FL2D, PSMD6, ABRX2, SART3, CND1, EXOS7, U5S1, SYLM, WDR43, ACOX1, EBP, PCM1, NOMO1, PON2, TEBP, NONO, RAB35, RNPS1, PCBP2, ELOC, UBE3C, DHC24, SF3B3, RSU1, SF3B4, SF3A1, SKI2, MARE2, NHRF2, TSN, SF01, CIP4, TRIPB, PCH2, ELAV1, SMAD2, TBCE, VAMP3, ZYX, SEPT7, CCDC6, UAP1, PSMD5, MK14, RM23, DPYL2, SYPL1, RBBP7, IF16, KYNU, CAH9, NDUA9, PCKGM, HCDH, UGPA, MIC60, HNRL2, INF2, PDS5A, TSR1, ALG11, QRIC1, SMU1, LRRF1, RPA43, LSM12, ACSF3, PPCEL, TB10B, MIX23, RHG29, PDCD4, Q53FH6, DHB12, SNUT2, LACB2, EIPR1, H90B4, TM41B, HERC4, PP6R3, PREP, PSMG4, RRP12, TOIP1, SYAM, PCID2, S27A3, SAMD9, CYTSB, TDIF2, TTC38, EXOS6, NU188, CE170, SNPC4, CHCH9, DDX59, GPTC4, HECD3, SKT, RM02, UBAP2, LCE1C, ACBD5, EEIG2, RBM26, DCAF8, ATPF1, DDI2, NT5D1, LYRM7, EMC10, RIF1, OBSCN, CE350, AXA81, PR38B, BRE1A, ARHGG, LYPL1, ECM29, MIC13, MAP1S, UTP25, TENS3, INT3, C19L1, CPIN1, SMYD5, PTRD1, CNDH2, MRM1, P4R3A, ACD10, DE10B, SCMC1, TM214, HIBCH, TTI2, DHX57, CDC73, RM14, L2GL2, MET2B, PRP8, TTC27, SKI3, RICTR, LCLT1, WDR82, HACD2, GGYF2, KAT3, UBR3, LARP4, SPT6H, SND1, COX15, DDX46, TM10C, 5MP2, EIF3M, MEPCE, CYFP1, ENOF1, FAKD5, MOB1B, KDM3B, HS2ST, K2C74, TRM1L, ZCCHV, NUP54, MAVS, DHX29, COMD6, HDDC2, LIMS2, I2BP2, PATL1, SETD3, PABP2, MTA70, ZN598, AACS, LUZP1, GP180, CAND1, CMTD1, PRSR2, CARM1, COMD2, DDX42, SPAS2, CONA1, NDUAB, DAAF5, STX12, DLP1, P66A, HORN, RB6I2, FABD, CEP97, CTL2, PHF6, UBR1, CHERP, ANKH1, DJC10, NELFD, MIRO1, RB12B, RAVR1, SPB1, SRRM1, CA174, MICU2, TRM2A, PELP1, ABI1, CCAR2, NDUF2, NUP93, LRC47, GPD1L, TOM5, OGFD1, TTC9C, JAGN1, CPSF7, PAF1, ZN433, F241A, ENAH, CL029, NHLC2, SUMF2, UBAC2, RDH13, UXS1, FA98A, NNRE, LS14A, PAPD5, CNNM3, MCU, FBX22, FAD1, NUP35, Q8NFH9, ABHDB, KNL1, OR1M1, ATLA2, MSPD2, DDX55, NCOA7, THOC2, WDR36, G45IP, CX038, FBX30, S35B2, PUM2, KLDC4, UBA3, KISHA, TBC15, RM30, PNPT1, SSH3, GEMI5, IPO4, ABHD5, ZC3HF, TF3C2, TM263, PTPM1, NU133, CHAC2, THEM6, LEO1, CPLX3, NUDC2, TFB1M, SCFD1, HNRLL, CD003, SEN15, SPRY4, TRUB1, LMO7, ARAP3, PRP31, PALLD, CTBL1, DDB2, DDX1, DHB8, PSMF1, RTF1, RT27, AP3S1, LAR4B, GCN1, PRP16, NU205, TFG, RBP56, DDX17, RAD50, CELF1, OSTF1, ARHG1, UFD1, RENT1, SMRC1, FUBP2, ARHG2, UBP13, USP9X, SCAM4, OSBP2, F162A, RM24, SYAP1, EXOC4, FERM2, PBIP1, RPE, EXOS8, OTUL, CHCH1, PPWD1, COA7, TAM41, F136A, P5CR2, DCPS, FAF2, FXRD1, AP2M1, GCP3, F241B, NMD3, RMD1, MSI2H, SNR40, RM38, RRFM, SIR1, YIPF6, KBP, GNA1, RT24, MOCOS, DAZP1, SAAL1, SIM12, DNJA3, TMA16, PTCD3, OTUB1, TRM61, HMCES, DUS3L, OTUD5, VMP1, DCNL1, SMRD1, TOE1, DDX27, MTNB, ZC21A, PGAM5, FUBP3, SPF45, SUCB2, HPDL, ATPMK, BT3L4, CLP1L, SCYL1, SNX27, YTDC1, ZN830, NSUN5, RBM14, UHRF2, PUS7, TRNT1, TEFM, EFGM, NUDC1, PRPK, PIGS, WRIP1, RANB9, NEB2, IWS1, SIN3A, OSBL9, INT14, ZSC10, RUFY1, MMS19, UHRF1, NIBA2, YMEL1, MYCBP, TBCB, PSB7, CNN2, CDC5L, PSMD1, PFD5, MPH6, TSNAX, TTC1, CSN8, KIF2C, MGLL, HCD2, NP1L4, ACON, TCPH, ANM1, VRK1, CND3, ARP5L, DDX50, RNZ2, GRWD1, MACD1, CMS1, MEP50, RT63, TBA1C, MBB1A, FERM1, CG050, PDD2L, MIC25, UQCC2, UTP23, LXN, NTPCR, TACO1, GCP2, SEN34, ALG1, PSMG3, CSN4, DIDO1, RBM42, MCMBP, AASD1, DCNL5, AN32E, P4K2A, MMTA2, TBB6, PAXX, HNRL1, MTNA, WDR18, THUM3, NTM1A, ASHWN, TM109, GNL3, NUP85, ELOV1, IFT27, SYTM, THIC, FUND2, SF3B5, SFXN3, CARD6, OSB10, B2L13, SRRT, HDHD5, HDAC8, RTF2, EIF2A, PDIP3, RM13, RM04, RM01, MK67I, YTHD1, RT26, NEUL, ABCA2, RM37, UACA, WDR11, CRNL1, TBL1R, UBL5, API5, FTO, UBE2O, XPO4, S39A8, WDR12, YIPF3, DERL2, SKI8, SLIRP, EGLN1, UBA5, MKRN2, T126A, PAIP1, CLPB, LSG1, NAT10, ILKAP, XRN2, NSRP1, CSTFT, 5NT3A, GBRL1, DDX47, RM18, SNAB, ATG5, GAN, DPH5, DHX36, IPYR2, SPNS1, RM46, PININ, GPT, GHITM, NELFA, OSGL1, HEAT1, TFB2M, YTDC2, RPAP3, WDR26, CH033, UBE2Z, ANKZ1, ZWILC, MED20, ARMT1, L2HDH, ELP3, RPA2, KT3K, SYCM, PPCS, NJMU, XPO5, GRPE1, RHOF, POPD2, EMAL4, GLOD4, MCCB, MOV10, SYF1, TM9S3, WDR6, TRXR2, SYSM, RT30, M1IP1, UBE2T, NOP10, OSGEP, MED4, RM40, RTN4, RPR1B, RRAGD, PFD4, EXOS3, ANLN, XPP1, BIRC6, ACSA, DBLOH, DDX21, SAR1A, DPOE4, SIAS, SHLB2, EI2BG, MBNL1, PYRG2, DPOE3, CHRC1, COXM2, ABC3C, PNO1, RM17, HELLS, SYFB, SYIM, ATG3, SMC4, OLA1, CUTC, ECHD1, RBM12, MDN1, DECR2, ABHDA, LIN7C, TXLNG, SPS2L, ABCF3, DD19A, UQCC1, EXD2, DJC11, FANCI, ATD3A, ARL8B, ANM7, FAIM1, RT18A, ARMC1, RPC5, SDA1, RBM28, RBM22, DMAC2, WDR70, PK1IP, RM22, CZIB, BABA1, NDUBB, RM16, NHP2, OCAD1, ADPRS, LYAR, MIC19, THUM1, TRM1, PP4R2, DPP3, BCLF1, FKB11, TMOD3, ELOV5, GTSE1, TECR, ERAP1, ARP10, CISD1, F120A, IF2B1, COQ3, HPBP1, MAT2B, CNOT2, OGFR, CNIH4, RM15, THYN1, KCMF1, TM14C, HM20B, RCC2, RELCH, BCCIP, STK26, FPRP, IBTK, RRBP1, SYLC, SUCB1, DPM3, ATX10, TFP11, SAE1, COMD3, MTRR, ASH2L, DHCR7, EIF3K, SAE2, WNT16, CSN7A, QCR9, ZO2, LRWD1, ABCF2, SYWM, SEC63, SRP68, CHRD1, BAP29, S30BP, PFD2, GPN3, PUF60, NRBP, ENOPH, NDUAC, EI2BD, VATH, XPO7, VPS51, S2513, DCTN4, FBX2, CCNL1, DBR1, FEM1B, DJC12, MRT4, CPSF3, NUDT5, RCOR1, TF3C4, PACN3, ACINU, AGO2, PSME2, PALD, COR1C, AKP8L, ALK, DD19B, PRP19, UBQL1, SNX12, CHIP, PACN2, MAGD2, SNX6, SSRG, FAF1, PPIE, MINP1, SCAF8, TRI33, PHF8, LIMC1, UBP24, SMC3, SCML2, PIN4, PSF2, EIF3L, PLAP, RUVB1, BCS1, SYFA, CNPY2, PKHA6, RRP44, RT07, PDIP2, WBP11, NOP58, SGT1, ACOT9, AAR2, MEMO1, TMX2, LC7L2, DHRS7, RT02, SBDS, TMED5, EXOS1, SF3B6, RM11, RRP15, NOP16, TPRKB, PPIL1, CIA2B, RT16, RT23, PTH2, RTCB, FBX7, NOC2L, RL36, SAMH1, HBS1L, DCAF1, UBP15, TBL2, TELO2, LAS1L, AFG32, ARI1, SAM50, PRC2C, PPME1, AP4S1, YTHD2, CTDP1, SP16H, UTP18, TNPO3, GMPPB, RBM8A, SNX8, FHOD1, SERC, NCOR2, ORNT1, ALG5, RT18B, CLIC4, NFS1, SPCS1, MD1L1, STK24, 5MP1, COMDA, DC1L1, TX264, S4A7, NDUB9, MORC2, S23IP
Species: Homo sapiens
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Zhao P, Viner R, Teo CF, Boons GJ, Horn D, Wells L. Combining high-energy C-trap dissociation and electron transfer dissociation for protein O-GlcNAc modification site assignment. Journal of proteome research 2011 10(9) 21740066
Abstract:
Mass spectrometry-based studies of proteins that are post-translationally modified by O-linked β-N-acetylglucosamine (O-GlcNAc) are challenged in effectively identifying the sites of modification while simultaneously sequencing the peptides. Here we tested the hypothesis that a combination of high-energy C-trap dissociation (HCD) and electron transfer dissociation (ETD) could specifically target the O-GlcNAc modified peptides and elucidate the amino acid sequence while preserving the attached GlcNAc residue for accurate site assignment. By taking advantage of the recently characterized O-GlcNAc-specific IgG monoclonal antibodies and the combination of HCD and ETD fragmentation techniques, O-GlcNAc modified proteins were enriched from HEK293T cells and subsequently characterized using the LTQ Orbitrap Velos ETD (Thermo Fisher Scientific) mass spectrometer. In our data set, 83 sites of O-GlcNAc modification are reported with high confidence confirming that the HCD/ETD combined approach is amenable to the detection and site assignment of O-GlcNAc modified peptides. Realizing HCD triggered ETD fragmentation on a linear ion trap/Orbitrap platform for more in-depth analysis and application of this technique to other post-translationally modified proteins are currently underway. Furthermore, this report illustrates that the O-GlcNAc transferase appears to demonstrate promiscuity with regards to the hydroxyl-containing amino acid modified in short stretches of primary sequence of the glycosylated polypeptides.
O-GlcNAc proteins:
P121C, MCRI1, ARI1A, TCRG1, MYPT1, HNRDL, SET1A, OGT1, HNRPR, ZN207, DNJA2, ANR17, EIF3G, EIF3J, SC31A, SC24B, TXD12, TBB4A, G3P, S10A8, RLA2, ENOA, NPM, PROF1, SYEP, HNRPC, SP1, VIME, GSTP1, ROA1, H14, THIO, HSP7C, PABP1, ENOB, PO2F1, HNRPL, RS2, RL17, NUCL, RO52, PIMT, SFPQ, EF1B, RS12, DDX6, RL13, ERP29, RL12, PPIF, DNJA1, HSP74, RL22, FUS, NU214, SRP14, NUP62, RBMX, GRP75, RANG, RL27A, RL5, RL21, RL28, RL29, RL34, SRP09, NU153, TCPD, HCFC1, ROA3, AGFG1, NUP98, SC24C, SEC13, TPIS, RS20, RL26, WDR5, RS7, RS14, RS23, RS11, RL23A, RS6, RAN, RL23, RL31, RL11, RL8, PPIA, YBOX1, EF1A1, TBA1B, RAE1L, RL36A, HNRPU, RL18A, KMT2A, C1QBP, KHDR1, AIMP1, ILF3, ROA0, CBX3, CIRBP, VEZF1, UBP2L, NOLC1, SYK, RBBP5, SC23A, SF01, JHD2C, ELAV1, QSER1, LSM12, SNUT2, UBAP2, RBM26, LIN54, NFRKB, NUP54, NUFP2, EMSY, CARM1, ANKH1, CCAR1, SACA3, ASXL1, BAP18, SERB1, FA98A, SMRC2, ATX2L, DDX1, HS105, TFG, DDX17, SMRC1, FUBP2, ZC3HA, CIC, NUP88, ATX2, ROAA, DIDO1, NUP58, WNK3, TBL1R, WNK1, DMAP1, DDX21, BCLF1, CDK12, KANL3, ASH2L, NUP50, YETS2, RTRAF, RUVB2, RUVB1, AT8A1, STRAP, RTCB, WNK2, PRC2C, S23IP
Species: Homo sapiens
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