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Liu J, Hao Y, Wang C, Jin Y, Yang Y, Gu J, Chen X. An Optimized Isotopic Photocleavable Tagging Strategy for Site-Specific and Quantitative Profiling of Protein O-GlcNAcylation in Colorectal Cancer Metastasis. ACS chemical biology 2022 35254053
Abstract:
O-linked-β-N-acetylglucosamine (O-GlcNAc) glycosylation is a ubiquitous protein post-translational modification of the emerging importance in metazoans. Of the thousands of O-GlcNAcylated proteins identified, many carry multiple modification sites with varied stoichiometry. To better match the scale of O-GlcNAc sites and their dynamic nature, we herein report an optimized strategy, termed isotopic photocleavable tagging for O-GlcNAc profiling (isoPTOP), which enables quantitative and site-specific profiling of O-GlcNAcylation with excellent specificity and sensitivity. In HeLa cells, ∼1500 O-GlcNAcylation sites were identified with the optimized procedures, which led to quantification of ∼1000 O-GlcNAcylation sites with isoPTOP. Furthermore, we apply isoPTOP to probe the O-GlcNAcylation dynamics in a pair of colorectal cancer (CRC) cell lines, SW480 and SW620 cells, which represent primary carcinoma and metastatic cells, representatively. The stoichiometric differences of 625 O-GlcNAcylation sites are quantified. Of these quantified sites, many occur on important regulators involved in tumor progression and metastasis. Our results provide a valuable database for understanding the functional role of O-GlcNAc in CRC. IsoPTOP should be applicable for investigating O-GlcNAcylation dynamics in various pathophysiological processes.
O-GlcNAc proteins:
A0A0B4J203, A0A0C4DFX4, RBM47, E2F8, WDR27, SBNO1, CNOT1, P121B, P121C, H0YAE9, H0YHG0, H7C469, K7ELQ4, M0QZ24, PDLI1, HAX1, TAF4, BCL9, CAC1A, DDX3X, NFIB, PPP6, MA2B1, ARI1A, SOCS7, ABLM1, KMT2D, GBRD, RGRF2, TX1B3, HGS, MYPT1, SYN3, ZN609, TRI66, PDZD2, MAST4, SC16A, SET1A, CASC3, FOXP2, MOT4, P4HA2, ARPC5, CLOCK, MAFG, PER1, KDM6A, TET3, SI1L1, TGFI1, M3K7, MCA3, PRPF3, TPD54, SYNJ1, IF4G3, E41L2, WIPF1, FOXO3, TGM5, RNF13, SPY2, PLRG1, ZN207, AKAP8, CALU, ORC5, MYPT2, GANP, OGA, CCNT1, BUB1B, PLOD3, PLIN3, MOT2, MAFK, PQBP1, BRD4, TBL1X, PP1RB, NBN, MITF, SRGP2, N4BP1, ROCK2, PP6R2, CNOT3, ANR17, FLNB, NCOR1, SF3B1, REM1, CREG1, CRTAP, SYUG, CYTF, TOX4, TOX, SUN1, PCF11, AGFG2, UBE4B, CAC1H, SVIL, SC24A, SC24B, CNOT4, EYA4, ZMYM6, BAG3, LATS1, DDAH2, TXD12, ONEC2, CLPT1, ABL1, CRYAB, LMNA, TFR1, CATA, GLCM, FUCO, ALDOA, GCR, G3P, CPNS1, HSPB1, RLA2, RLA0, ITB1, K1C18, NPM, CATL1, CATB, MCR, BGLR, ITA5, NFIC, VIME, SNRPA, FGR, ATX1L, DERPC, ZN865, GLI2, MYBB, CLUS, PPAL, MPRI, PABP1, TPR, BMP3, SKIL, ENPL, PO2F1, PLAK, ATF2, ZEP1, RS2, TFE2, F261, ITB4, ZNF23, ZNF25, JUNB, ATF7, TPH1, DDX5, EGR1, SON, NELFE, ATF1, ATF6A, CADH2, ICAL, CSRP1, FLNA, RFX1, CBL, SFPQ, COF1, IF4B, GATA2, APC, DDX6, ARNT, MAP4, LYOX, HXD9, MZF1, CLIP1, 5HT1F, HXA11, ZEP2, ELF1, CTNB1, FBN1, ADDA, BASI, NU214, VGFR2, SRP14, NUP62, SYUA, VATA, CUX1, TXLNA, STAT3, LAP2A, EPS15, HELZ, MATR3, SSRA, SSRB, KI67, ATRX, MAP1B, YAP1, UTRN, STT3A, SC6A8, RFX5, SOX2, PRC2A, HSP13, NR2C2, NASP, CDK8, DHE4, YLPM1, NU153, RBP2, TAF6, MRE11, EMD, MXI1, MAP2, TOB1, PPT1, TCPQ, PAPOA, HCFC1, GDS1, AGFG1, CRIP2, NUP98, SMTN, SC24C, HIRA, ATX1, ATN1, AFAD, AF10, AF17, DSRAD, SEC13, NU107, ZN445, CSN2, RL37, WDR5, TIM10, F193A, RBM6, PITX1, IF4G2, PHC1, ADA17, KGD4, RL19, FOXK1, DAB2, RHG04, RBM10, HNRPU, SPTB2, FOXK2, RUNX1, MEF2A, SP2, SP3, PLOD1, KMT2A, TF65, IF4G1, NOTC2, TLE3, TLE4, PTN12, CALD1, MEF2C, P5F1B, GABPA, ZO1, ACK1, EP300, AHNK, FCHO2, HMGX3, SRBP2, FOXO1, ASPH, TROAP, BPTF, FSTL1, NFIA, DPYD, TP53B, FOXC1, ECH1, ROA0, DDX10, TBX2, GPS2, G3BP1, PABP4, ADAM9, PICAL, NAB1, SERC3, RIPK1, IQGA2, STIM1, CUL4B, ASPP2, CAC1S, RUNX2, NFYC, CDK13, TOB2, VEZF1, UBP2L, GIT2, SRC8, CAPR1, LAGE3, PUM1, MDC1, EPN4, TTLL4, RRP1B, NCOA6, GSE1, MEF2D, LASP1, MYPC3, ZN638, NUMA1, SART3, CND1, R3HD1, KIF14, WDR43, PLCL1, PLEC, NOMO1, NONO, RCN1, RYR3, KS6A1, RBMS2, TAF1C, SF01, MED1, JHD2C, TRIP6, T22D1, ELF2, TAB1, HERC1, NCOA1, VAS1, ZFHX3, ZYX, ADRM1, SYPL1, TAF9, DREB, DGKD, CGT, GEN, LY6K, RFX7, QSER1, AAK1, PRSR3, QRIC1, MA7D1, WDR72, TBRG1, TB10B, TPRN, FIL1L, SVEP1, AMOT, EPC2, CRTC2, PAN3, HS904, YIF1B, AG10A, IGS11, ZN628, BCORL, FIGN, K2026, SH319, TGO1, PRC2B, TOIP1, CEP78, P4R3B, HP1B3, CE170, ZN362, FKB15, AKND1, ZEP3, LRIF1, SWT1, RHG21, UBAP2, RBM26, DEP1A, OGRL1, AHDC1, F222A, RPRD2, RN220, ZN318, TASO2, ZMYM4, PAPD7, TNS2, KANK2, ARID2, USF3, RHG17, CYTSA, ANR40, BICRL, JADE1, PKHA7, NIPBL, LIN54, TET2, RINT1, CRCDL, ZNT6, TTC41, RHGBA, NFRKB, RSBNL, KCD18, NCEH1, MDEAS, ZC3HE, LARP1, NHS, CRTC3, SAS6, MCAF1, BCOR, MPRIP, DNMBP, GGYF2, THADA, BNC2, NFXL1, NBEL2, CO039, SRCAP, CBAR2, UBN2, XIRP1, RAPH1, LARP4, HAKAI, ASXL2, SPT6H, KDM3B, ZCCHV, KANL1, RGPD4, POGZ, ZFY16, NUFP2, MAVS, CLAP1, EMSY, I2BP2, SRGP1, RBBP6, SH3R1, HUWE1, YTHD3, NPM2, ILDR1, KAISO, MYPN, LDB1, LYRIC, BCL9L, LUZP1, NRAP, RTTN, PRSR1, DDX42, CEP57, CD20B, CACL1, P66A, HIPK1, KCC1D, RN135, MY18B, AHNK2, FOXP4, NAV3, NAV2, MISP, ARI3B, IPRI, TEX2, MGAP, CC28A, Z3H7A, ANKH1, SUGP1, RPAP2, MILK2, SRRM1, ZZZ3, FA71A, PHAR4, RTKN2, DCP1B, XRN1, PELP1, CKLF8, TENS4, SPART, RPTOR, NUP93, ZN687, DOCK4, RHG24, RUSC2, SYNPO, FNBP4, D2HDH, RP25L, ATPF2, CPSF7, ARFG1, ENAH, SPOT1, SUMF1, KCNH5, SLAI1, TNR6A, PHC3, DRC6, CBPC3, NAV1, VP37A, KMT2C, ZMIZ2, BD1L1, ARI1B, FLCN, NUP35, TOIP2, TNIP2, KNL1, OR2L2, PUM2, CC110, TBC15, STT3B, ZN507, ALMS1, DLG5, KCNV2, BRX1, DOT1L, GEMI5, PARD3, ZN384, SMAP2, IASPP, TM263, ZFN2B, NUDC2, PCNP, TRUB1, LMO7, ATX2L, PALLD, P66B, BBX, ZCH14, GBF1, SMG7, RTF1, NICA, PHF3, MAML1, ZN592, LAR4B, TFG, TAF4B, RREB1, SC65, CBP, SYMPK, DDX17, GPKOW, FUBP2, UBP7, LPP, LSM10, NCLN, MRTFA, FUBP1, TTC17, PBIP1, TTC28, TOM6, PF21A, INT12, REPS1, ESS2, MBD6, ELP4, SGF29, RBM33, ZN503, P121A, TONSL, PDLI5, ERO1A, DOCK6, FUBP3, RSRC1, ZN594, VCIP1, ZN462, LCOR, PDLI2, CLP1L, Z512B, ZFR, EP400, MRFL, H6ST2, TIGD1, NOL4L, DOCK7, RPR1A, RBM14, ADCYA, QKI, LENG8, TRNT1, PP1RA, PHF12, CIC, MED15, ERBIN, HMCN1, LMF1, PIGS, WRIP1, SIN3A, MINT, HTF4, EYA3, POP1, TEAD3, TTC1, CSN8, ATX2, ARI3A, ANM1, PKP2, TEP1, DPH2, WAC, DIDO1, HNRL1, RBM4, SSBP4, PRR14, SSBP3, YTHD1, KPCD2, ZCHC2, TB182, AMRA1, CE295, TANC1, ZC12C, CEP44, STRAB, SP130, BRD8, RGAP1, SMG9, APC1, I2BPL, TMX4, KI13A, WDR13, EPC1, ADNP, ZN106, TM245, FOXP1, PABP3, WNK1, E41L1, ZHX3, BICC1, PEAK1, PPR3E, ZN703, PKHA5, CLSPN, BCDO1, RC3H2, ZFYV1, TAF9B, EMAL4, ZBT20, NCOA5, TANC2, ZN532, NCK5L, TNR6C, CHD8, FBSL, APMAP, DMAP1, UBN1, DCP1A, INCE, ANLN, GEPH, PDLI7, TULP4, HOME2, SLX9, DIAP3, BMP2K, RBM12, STAU2, DDX28, CWC25, CARF, ETAA1, ABI2, HXC10, BCLF1, TAB2, CELR3, CDK12, GRHL1, SACS, ITSN2, BICRA, CNOT2, TMEM9, CAC1I, CAMP3, DAPLE, RCC2, DIP2B, MBD5, CT2NL, F135A, KANL3, RERE, SE1L1, TRM7, YM012, KDM5B, LIMD1, TCF20, SUN2, LIMA1, SEPT9, UBQL2, TRPS1, S30BP, NRBP, BAZ2B, SIX4, HOOK1, CDC23, TASOR, GMEB2, TNIK, PARP4, NUP50, ZHX1, CDV3, MCTS1, KCNH3, LRFN2, MRTFB, ZBT21, PRR12, YETS2, HECD1, PKCB1, NOTC3, SPAT2, SOX13, G3BP2, MAGD2, MINP1, MACF1, CP131, SCAF8, TRI33, PHF8, LIMC1, TNR6B, SRRM2, SCML2, ZN148, POLH, INVS, ICE1, R3HD2, MAN1, TR150, WBP11, ZN281, STA13, WNK2, HBS1L, ARIP4, MTCL1, DCAF1, RPGF2, IRS2, CRBG1, HYOU1, SAM50, PRC2C, YTHD2, NCOR2, GMEB1, DC1L1, EPN1, NCOA3, ZHX2, S23IP, U3KPZ7, V9GYH0
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 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, GLCM, 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, SL7A1, 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, MPCP, 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, TTC37, NCEH1, BRAT1, ZC3HE, C1TM, SBSN, CREL2, PACS1, GGYF2, SRCAP, TMTC3, IKIP, SPT6H, SND1, DDX46, BZW1, 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, PAIRB, 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, PKCB1, 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, TRUA, NPTN, S12A7, COPG1, BIG1, BZW2, DC1L1, IF2B2, ZNT1, S4A7, ZHX2, S23IP
Species: Homo sapiens
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Li J, Li Z, Duan X, Qin K, Dang L, Sun S, Cai L, Hsieh-Wilson LC, Wu L, Yi W. An Isotope-Coded Photocleavable Probe for Quantitative Profiling of Protein O-GlcNAcylation. ACS chemical biology 2019 14(1) 30620550
Abstract:
O-linked N-acetylglucosamine ( O-GlcNAc) is a ubiquitous post-translational modification of proteins and is essential for cell function. Quantifying the dynamics of O-GlcNAcylation in a proteome-wide level is critical for uncovering cellular mechanisms and functional roles of O-GlcNAcylation in cells. Here, we develop an isotope-coded photocleavable probe for profiling protein O-GlcNAcylation dynamics using quantitative mass spectrometry-based proteomics. This probe enables selective tagging and isotopic labeling of O-GlcNAcylated proteins in one step from complex cellular mixtures. We demonstrate the application of the probe to quantitatively profile O-GlcNAcylation sites in 293T cells upon chemical induction of O-GlcNAc levels. We further applied the probe to quantitatively analyze the stoichiometry of O-GlcNAcylation between sorafenib-sensitive and sorafenib-resistant liver cancer cells, which lays the foundation for mechanistic investigation of O-GlcNAcylation in regulating cancer chemoresistance. Thus, this probe provides a powerful tool to profile O-GlcNAcylation dynamics in cells.