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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.
O-GlcNAc proteins:
A0A0B4J203, A0A0C4DFX4, SBNO1, P121B, CX028, RGPD3, AN36A, P121C, GG6L6, S31C2, E9PCH4, H0YAE9, H0YHG0, GG6LV, H3BMH7, PDLI1, TAF4, BCL9, ABLM1, CHD2, KMT2D, RGPD8, OPLA, HGS, MYPT1, ZN609, SC16A, SET1A, TIF1A, EIF3H, TET3, M3K7, PRPF3, TPD54, IF4G3, E41L2, AKAP8, TM11D, MYPT2, GANP, PLIN3, MAFK, BRD4, MITF, N4BP1, ATG13, PP6R2, ANR17, NCOR1, SPAG7, SRS10, SF3B1, CSDE1, TOX4, PCF11, AGFG2, SMC2, M3K6, SC24B, ZBT11, CNOT4, EYA4, OXSR1, ZMYM6, CCNE2, ANGT, LMNA, ALDOA, GCR, HSPB1, F13B, RLA2, K1C18, K2C8, ZFY, SRPRA, RU17, VIME, RU2A, ATX1L, RGPD1, S31C1, GLI3, LYAG, PABP1, COBA1, CO6A3, MYH7, ENPL, ZEP1, RS2, ZFX, ZNF30, ANPRC, ATF7, EGR1, SON, RCC1, ATF1, ATF6A, HXA5, ROA2, CBL, IF4B, GATA2, RIR1, RAE1, APC, ATPA, ARNT, MAP4, HXD9, HLAF, CLIP1, ZEP2, MYH10, TIE1, NU214, DEK, PDE6B, SRP14, CUX1, LPPRC, GATA4, KI67, YAP1, RFX5, SOX2, PRC2A, NASP, CDK8, NU153, RBP2, TAF6, EMD, PAPOA, NEK4, AGFG1, NUP98, INHBC, CADH6, F193A, KGD4, RT34, SARNP, LACTB, COG7, FOXK1, DAB2, PLIN5, SPTB2, SP2, NRG1, IF4G1, K1C17, TLE1, TLE4, UBE3A, ACK1, AHNK, FCHO2, FOXO1, TROAP, BPTF, IRAG2, BFSP1, FOXC1, PRDM2, DDX10, G3BP1, PABP4, GRB10, PPIG, MADCA, PICAL, MAMD1, CUL4B, ASPP2, SPTN1, CDK13, CYLC2, DSG2, UBP2L, SRC8, ITPR3, PUM1, MDC1, EPN4, RRP1B, NCOA6, RRP5, RFTN1, R3HD1, WDR43, EEA1, MTFR1, SF3B3, RYR3, SF01, JHD2C, ELF2, MYLK, TAB1, ZYX, ADRM1, QSER1, CL16A, RHG31, AAK1, TMM44, AMOT, IF44L, YIF1B, AG10A, CD048, FSIP2, ESCO1, S2553, BCORL, AN36C, MTUS2, PRC2B, CEP78, SAMD9, TSBP1, LRIF1, CXG2, SKT, ZN648, UBAP2, RBM26, RC3H1, EFCB6, CE350, RPRD2, S31A6, TASO2, ECM29, RN123, PLCX3, ARID2, DEN2C, K0930, LIN54, M18BP, SCYL2, NFRKB, KLH35, ZC3HE, ANR11, FIP1, SBSN, S49A3, FAT4, MCAF1, BCOR, DUSTY, GGYF2, BNC2, CO039, SRCAP, UBN2, FOXNB, UBP54, HAKAI, ASXL2, KNDC1, SPT6H, TAOK1, KDM3B, RGPD4, POGZ, NUFP2, EMSY, I2BP2, SH3R1, HUWE1, YTHD3, FLIP1, KAISO, MYPN, TTC6, LDB1, TM135, TBC26, ZFHX4, ANGL5, SPAS2, DZIP1, P66A, AHNK2, FMNL3, NAV2, ARI3B, MGAP, RP1L1, CC28A, Z3H7A, CDAN1, ANKH1, SUGP1, PHAR4, KMT2E, XRN1, SPART, NUP93, ZN687, CMTR1, THMS1, AN36B, TMTC2, SYNPO, FNBP4, GG6L1, ENAH, GG6L2, SLAI1, PHC3, BD1L1, NUP35, DDX55, NEIL3, GSDMB, ALMS1, STK35, GEMI5, RPGF6, SMCR8, WIPF2, TM171, RN133, TEKT4, LMO7, CKAP2, ATX2L, ACO11, P66B, DAAF4, BBX, FIG4, ZN516, RREB1, FUBP2, LPP, E2F7, TTC28, TOM6, ASTRA, OTUB2, PGBD4, LEG12, ELP4, RBM33, MYEOV, SMRD1, DDX27, P121A, TONSL, PDLI5, THOC3, VCIP1, LRIQ1, ZFR, EP400, CBPC4, RBM14, IPP2L, QKI, PLIN4, JMJD8, RBM15, MINT, SEC62, AGAP2, RGPD5, ATX2, MYD88, ARI3A, SPI2A, SPI2B, DPH2, MCMBP, TMPSD, YTHD1, WNK3, PP12C, TB182, TANC1, CEP44, SENP6, BRD8, RGAP1, ALX4, KI13A, KCNH6, ZN106, FOXP1, PABP3, SMOC2, WNK1, ZHX3, CP095, REEP4, DOCK5, ZN703, GORS2, MLXIP, PKHA5, FOH1B, RC3H2, TANC2, TRPM3, SYTL2, CP4FC, GAK5, JPH1, APMAP, DMAP1, GP108, KMT5A, GPR84, DUOX2, DUOX1, PCDBG, MDN1, NALP2, CARF, HXC10, TAB2, CDK12, ADA2, ITSN2, F135A, SI1L2, RBM27, KANL3, ZN219, DYH17, AFF4, NB5R1, S30BP, NRBP, BAZ2A, SIX4, HOOK1, TASOR, GMEB2, ZHX1, TAOK2, CFA92, MRTFB, ZBT21, PRR12, YETS2, HECD1, MYO6, ICAM5, MAGD2, SCAF8, TRAK1, SHAN2, SRRM2, EXO1, SCML2, POK19, POLH, NCKP1, AT11B, NOP58, ZN281, UB2J1, GRIP1, SALL2, ARIP4, RPGF2, HYOU1, TTLL3, PRC2C, PCDB4, NCOR2, CP46A, 5MP1, CABIN, NCOA3, S23IP, U3KPZ7
Species: Homo sapiens
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Qin W, Qin K, Fan X, Peng L, Hong W, Zhu Y, Lv P, Du Y, Huang R, Han M, Cheng B, Liu Y, Zhou W, Wang C, Chen X. Artificial Cysteine S-Glycosylation Induced by Per-O-Acetylated Unnatural Monosaccharides during Metabolic Glycan Labeling. Angewandte Chemie (International ed. in English) 2018 57(7) 29237092
Abstract:
The unexpected, non-enzymatic S-glycosylation of cysteine residues in various proteins by per-O-acetylated monosaccharides is described. This artificial S-glycosylation greatly compromises the specificity and validity of metabolic glycan labeling in living cells by per-O-acetylated azido and alkynyl sugars, which has been overlooked in the field for decades. It is demonstrated that the use of unacetylated unnatural sugars can avoid the artifact formation and a corrected list of O-GlcNAcylated proteins and O-GlcNAc sites in HeLa cells has been assembled by using N-azidoacetylgalactosamine (GalNAz).
O-GlcNAc proteins:
RBM47, SBNO1, CNOT1, PDLI1, TAF4, AGRIN, ABLM1, CSKP, MYPT1, ZN609, SC16A, NUP42, KDM6A, MSI1H, HNRPR, TPD54, SYNJ1, KLF4, RA51C, ZN207, NPA1P, DIAP1, PLIN3, MAFK, ANR17, LIPA2, NCOR1, SC24B, CNOT4, TXD12, EGFR, LMNA, GCR, HSPB1, RLA2, K1C18, VIME, ROS1, SNRPA, ROA1, IFIT2, ATX1L, SP9, TPR, MYH1, XRCC6, GLU2B, PO2F1, ATF2, ZEP1, RS2, ITB4, JUNB, UBF1, ATF7, DESM, SON, EFNA1, CSRP1, CBL, ARNT, MAP4, CLAT, MPIP1, CLIP1, NU214, NUP62, VKGC, VATA, CUX1, PEX19, LMAN1, NASP, CDK8, NU153, RBP2, TAF6, EMD, PAPOA, AGFG1, VAV2, NUP98, AF10, F193A, PITX1, KGD4, FOXK1, DAB2, HNRPU, SPTB2, IF4G1, TLE4, ZO1, AHNK, BPTF, TP53B, PTN13, G3BP1, OS9, PICAL, RIPK1, MTMR3, CUL4B, NCOA4, CDK13, VEZF1, DSG2, UBP2L, DYHC1, SRC8, DHX8, PUM1, MDC1, RRP1B, NCOA6, MEF2D, EBP, RBMS2, TAF1C, NCOA2, SF01, JHD2C, ELF2, TAB1, ZFHX3, ZYX, ADRM1, TAF9, QSER1, PRSR3, FBX43, EPC2, CRTC2, YIF1B, ESCO1, BCORL, TGO1, PRC2B, RHG21, UBAP2, RBM26, SPD2A, AHDC1, RPRD2, ZN318, CCD30, TASO2, ARID2, DEN2C, ANR40, BICRL, NIPBL, LIN54, NFRKB, MDEAS, ZC3HE, FIP1, CRTC3, SAS6, MCAF1, BCOR, GGYF2, RBM44, CO039, UBN2, HAKAI, ASXL2, SPT6H, KDM3B, ATG9A, MAVS, EMSY, I2BP2, SH3R1, HUWE1, YTHD3, OSTA, VS10L, CHD1L, PRSR1, DDX42, P66A, SAM9L, NAV2, TEX2, MGAP, ANKH1, SUGP1, SUGP2, CKP2L, SRRM1, CPNE9, XRN1, SPART, T132C, SYNPO, FNBP4, KASH5, ARFG1, ENAH, MLKL, TNR6A, PHC3, NAV1, VP37A, NUP35, NEDD1, AGR3, UBS3B, C2C4C, ZN384, WIPF2, TM263, LMO7, ATX2L, CSKI2, P66B, BBX, TITIN, SMG7, ZN592, LAR4B, CBP, GPKOW, LPP, TTC28, PF21A, ARL8A, ROP1L, RBM33, PDLI5, FUBP3, COG3, VCIP1, MY15B, Z512B, ZFR, EP400, DRC1, H6ST2, RBM14, LENG8, KI20B, CIC, MINT, NELL2, TEAD3, LYST, ATX2, DPH2, WAC, DIDO1, HNRL1, TIM21, T121B, GPR87, YTHD1, ABCA2, TANC1, SP130, F234A, I2BPL, SPTN4, ADNP, FOXP1, PTN23, WNK1, E41L1, ZHX3, ES8L2, MLXIP, PKHA5, RC3H2, TAF9B, SPON1, NCOA5, TANC2, ZN532, SYTL2, ZDBF2, UBN1, BIRC6, ARGL1, CARF, CELR3, CDK12, ITSN2, SELN, KCMF1, DAPLE, KANL3, VTI1B, TCF20, UBQL2, S30BP, TASOR, GMEB2, MUC12, ZHX1, PRR12, YETS2, HECD1, TRI33, LIMC1, ZC3H4, SRRM2, SCML2, AMOL2, RPGF2, PRC2C, SRPRB, NCOR2, S23IP
Species: Homo sapiens
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