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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,