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Wang J, Dou B, Zheng L, Cao W, Zeng X, Wen Y, Ma J, Li X. Synthesis of Na2S2O4 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, ACOC, 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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Hao Y, Fan X, Shi Y, Zhang C, Sun DE, Qin K, Qin W, Zhou W, Chen X. Next-generation unnatural monosaccharides reveal that ESRRB O-GlcNAcylation regulates pluripotency of mouse embryonic stem cells. Nature communications 2019 10(1) 31492838
Abstract:
Unnatural monosaccharides such as azidosugars that can be metabolically incorporated into cellular glycans are currently used as a major tool for glycan imaging and glycoproteomic profiling. As a common practice to enhance membrane permeability and cellular uptake, the unnatural sugars are per-O-acetylated, which, however, can induce a long-overlooked side reaction, non-enzymatic S-glycosylation. Herein, we develop 1,3-di-esterified N-azidoacetylgalactosamine (GalNAz) as next-generation chemical reporters for metabolic glycan labeling. Both 1,3-di-O-acetylated GalNAz (1,3-Ac2GalNAz) and 1,3-di-O-propionylated GalNAz (1,3-Pr2GalNAz) exhibit high efficiency for labeling protein O-GlcNAcylation with no artificial S-glycosylation. Applying 1,3-Pr2GalNAz in mouse embryonic stem cells (mESCs), we identify ESRRB, a critical transcription factor for pluripotency, as an O-GlcNAcylated protein. We show that ESRRB O-GlcNAcylation is important for mESC self-renewal and pluripotency. Mechanistically, ESRRB is O-GlcNAcylated by O-GlcNAc transferase at serine 25, which stabilizes ESRRB, promotes its transcription activity and facilitates its interactions with two master pluripotency regulators, OCT4 and NANOG.
O-GlcNAc proteins:
A0A087X1C1, A0A0B4J203, A0A0C4DFX4, SRCRL, SBNO1, CNOT1, IQCAL, RGPD3, LRIQ3, P121C, YU005, H0YHG0, H3BMQ9, H8Y6P7, I3L521, PDLI1, TAF4, P3C2A, DDX3X, NFIB, ARI1A, ABLM1, KMT2D, ZN197, RGPD8, MYPT1, ZN609, ZN646, SET1A, SYNEM, ZN185, NUP42, SI1L1, TGFI1, VIP2, M3K7, TPD54, SYNJ1, IF4G3, WIPF1, SMAD6, MYPT2, PLIN3, MAFK, N4BP1, ANR17, NCOR1, GGYF1, PRDM1, STAM2, TOX4, AGFG2, UNC5C, VENTX, SC24B, PCNT, ZBT11, CNOT4, BAG3, TXD12, TTLL1, APCL, NSD2, EGFR, IGF2, CRYAB, LMNA, ALDOA, GCR, HSPB1, PCCA, RLA2, JUN, HEP2, K1C18, ANXA2, ADRB2, NFIC, VIME, 5HT1A, SNRPA, ROA1, CO4B, ATX1L, FMAS1, GRL1A, DERPC, C1C1L, GLI2, TPR, GYS1, MYL6B, PO2F1, ATF2, ZEP1, RS2, ITB4, JUNB, JUND, ATF7, SON, ATF1, NEBU, CSRP1, NF1, ROA2, RFX1, CBL, COF1, IF4B, ARNT, MAP4, CALX, TEAD1, PDIA3, CDC27, CLIP1, ZEP2, ELF1, TTK, PCKGC, ADDA, NU214, MP2K2, NUP62, VKGC, VATA, CUX1, TXLNA, PBX2, HELZ, UTRN, RFX5, PAXI, NR2C2, NASP, CENPF, YLPM1, NU153, RBP2, TAF6, EMD, PAPOA, HCFC1, AGFG1, NUP98, ATX1, MYOM2, AF10, AF17, DSRAD, NU107, RL8, F193A, PITX1, PHC1, RT36, SARNP, FOXK1, HTD2, DAB2, RHG04, VIGLN, HNRPU, SPTB2, SCN7A, FOXK2, EWS, MEF2A, SP2, NUCB1, IF4G1, NOTC2, TLE3, TLE4, NMDZ1, GABPA, ZO1, ACK1, CACB2, EP300, AHNK, MGAT2, GALT2, FOXO1, SNF5, BPTF, NFIA, DPYD, TP53B, ZN155, FOXC1, AKAP6, ROA0, GPS2, G3BP1, KCAB2, PABP4, PLD1, PICAL, MAMD1, RIPK1, SNW1, MTMR1, CUL4B, ASPP2, NFYC, CDK13, TOB2, DAG1, VEZF1, DSG2, UBP2L, GIT2, SRC8, MDC1, EPN4, RRP1B, NCOA6, GSE1, MEF2D, ARI5B, NUMA1, PSME4, SART3, KIF14, BRD3, EBP, PLEC, RBMS2, TAF1C, SF01, MED1, JHD2C, MARE1, ELF2, TAB1, ZFHX3, ZYX, ADRM1, TAF9, RFX7, QSER1, QRIC1, ZN800, LR75B, EPC2, CRTC2, YIF1B, ZN326, K2026, PRC2B, SYAM, CE170, NHSL1, ZN362, ZEP3, LRIF1, UBR4, SKT, RHG21, UBAP2, RBM26, RC3H1, VP13D, RPRD2, ZN318, TASO2, TTC23, ERR2, KAZRN, ARID2, RHG17, ANR40, BICRL, NIPBL, LIN54, NFRKB, ZN449, RSBNL, MDEAS, ZC3HE, CRTC3, SAS6, MCAF1, BCOR, MPRIP, GGYF2, BNC2, FGD5, CO039, SRCAP, YJ005, UBN2, RAPH1, UBP54, PREX2, HAKAI, ASXL2, SPT6H, KDM3B, ZCCHV, RGPD4, POGZ, SZRD1, MAVS, EMSY, RAI1, I2BP2, SRGP1, SH3R1, YTHD3, STRA8, MYPN, TLK2, ZFHX4, BCL9L, IQCH, SNX32, PRSR1, MEX3D, DDX42, CACL1, P66A, CC125, KCC1D, HID1, AHNK2, FOXP4, NAV2, MGAP, RP1L1, PHF6, ANKH1, SUGP1, CCAR1, RPAP2, MILK2, EFC13, PHAR4, XRN1, TBC3A, TENS4, SPART, NUP93, ZN687, SYNPO, FNBP4, TAB3, CPSF7, ARFG1, ENAH, XXLT1, CHSTE, TNR6A, PHC3, SP20H, NAV1, VP37A, KMT2C, ARI1B, NUP35, TDRD7, NEDD1, PUM2, ALMS1, DLG5, ZN384, WIPF2, FRS2, F222B, SMAP2, IASPP, ZFN2B, TWST2, PCNP, LMO7, ATX2L, STAB2, PALLD, CSKI2, SRS12, P66B, BBX, SMG7, RTF1, PHF3, MAML1, LAR4B, PRP16, PRCC, CBP, EVPL, DDX17, GPKOW, FUBP2, LPP, FUBP1, TTC28, PF21A, KLH29, RBM33, MMAB, EF2KT, GWL, P121A, PDLI5, INT4, FUBP3, PAWR, ANCHR, Z512B, ZFR, EP400, COG8, RBM14, QKI, LENG8, CIC, MED15, ERBIN, MINT, HTF4, TEAD3, RGPD5, CGRE1, ATX2, CTIP, SH3G1, DPH2, WAC, DIDO1, TBCD, HNRL1, HIRP3, CAR11, YTHD1, GTPB4, AMRA1, TANC1, CEP44, MFF, SP130, BRD8, RGAP1, I2BPL, RBNS5, ADNP, FOXP1, PTN23, CA198, WNK1, E41L1, ZHX3, ILRUN, PEAK1, PKHG2, ECT2, JUPI2, PKHA5, RC3H2, TAF9B, ZBT20, NCOA5, TANC2, ZN532, ARFG3, PLPL8, UBN1, PCD12, INCE, PDLI7, 4ET, DIAP3, PDS5B, RBM12, CCD87, CARF, TAB2, MS18A, CDK12, ITSN2, WHRN, DAPLE, SLAI2, BAHC1, BCCIP, C2D2A, RBM27, KANL3, ZN219, LIMD1, TCF20, UBQL2, S30BP, NRBP, CTNA3, BAZ2B, HERC5, SIX4, TASOR, GMEB2, PARP4, NUP50, ZHX1, MRTFB, PRR12, YETS2, HECD1, SPAT2, SCAF8, LIMC1, ZC3H4, SRRM2, SCML2, ZN148, ZFP30, WDR37, MYH15, R3HD2, ZN281, COQ6, DCAF1, RPGF2, CRBG1, PRC2C, RBM7, CD2AP, TSSC4, HCFC2, NCOR2, GMEB1, DC1L1, NCOA3, ZHX2, S23IP, U3KPZ7
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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, CJ052, 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, HCFC1, AGFG1, VAV2, NUP98, AF10, F193A, PITX1, RT36, FOXK1, DAB2, HNRPU, SPTB2, IF4G1, TLE3, 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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Qin K, Zhu Y, Qin W, Gao J, Shao X, Wang YL, Zhou W, Wang C, Chen X. Quantitative Profiling of Protein O-GlcNAcylation Sites by an Isotope-Tagged Cleavable Linker. ACS chemical biology 2018 13(8) 30059200
Abstract:
Large-scale quantification of protein O-linked β- N-acetylglucosamine (O-GlcNAc) modification in a site-specific manner remains a key challenge in studying O-GlcNAc biology. Herein, we developed an isotope-tagged cleavable linker (isoTCL) strategy, which enabled isotopic labeling of O-GlcNAc through bioorthogonal conjugation of affinity tags. We demonstrated the application of the isoTCL in mapping and quantification of O-GlcNAcylation sites in HeLa cells. Furthermore, we investigated the O-GlcNAcylation sensitivity to the sugar donor by quantifying the levels of modification under different concentrations of the O-GlcNAc labeling probe in a site-specific manner. In addition, we applied isoTCL to compare the O-GlcNAcylation stoichiometry levels of more than 100 modification sites between placenta samples from male and female mice and confirmed site-specifically that female placenta has a higher O-GlcNAcylation than its male counterpart. The isoTCL platform provides a powerful tool for quantitative profiling of O-GlcNAc modification.
O-GlcNAc proteins:
A0A0A6YVU8, A0A1B0GSG7, RBM47, ZN335, A2A8N0, TITIN, SBNO1, CNOT1, PHRF1, ZN462, TAGAP, D3YUK0, E9PUR0, E9PVW1, E9PWI7, PARP4, E9PZS2, E9Q2C0, E9Q3G8, E9Q616, BD1L1, E9Q732, ARHG5, E9Q7N9, E9Q842, E9Q9B4, E9Q9Q2, E9QA22, E9QAE1, F6Y6L6, F8VQ29, F8VQM5, J9JI28, PDLI1, SPT5H, TAF4, ARI1A, ABLM1, KMT2D, MYPT1, ZN609, SET1A, SYNEM, PUR4, TNC18, KDM6A, DPOD2, M3K7, TPD54, SYNJ1, ZN207, SRPK2, ACK1, SYUA, MYPT2, KIF1B, HBP1, OGA, VINEX, PLIN3, MAFK, BRD4, PDLI1, KDM6A, SRPK1, N4BP1, ANR17, NCOR1, CREG1, CRTAP, MYO1A, MTR1L, CREG1, TOX4, SUN1, M3K6, PSMG1, SC24B, CNOT4, ABL1, ABL1, EGFR, LAMC1, LMNA, GLCM, GCR, HSPB1, PPBT, RLA2, ITB1, K1C18, K2C8, SAP, CATL1, LAMB1, ENPL, BGLR, NFIC, VIME, SNRPA, ROA1, ATX1L, TGAP1, GLI2, HLAC, CATB, TAU, BIP, FINC, K2C8, TPR, MSH3, ENPL, PO2F1, ATF2, GNS, ZEP1, RS2, MUC1, JUNB, ATF7, CATD, SON, SERPH, NELFE, BIP, ROA2, CBL, IF4B, APC, ARNT, MAP4, TEAD1, RXRA, RXRB, RXRG, CLIP1, AIMP1, HXA11, ELF1, NU214, MP2K2, VATA, CUX1, PBX2, MLH1, STAT3, SSRB, KI67, STT3A, RFX5, LMNA, DPOD2, PAXI, CDK8, YLPM1, NU153, RBP2, TAF6, EMD, PPT1, FXR1, ICAL, HCFC1, AGFG1, NUP98, ATX1, ATN1, PTN5, AF17, DSRAD, AMRP, ACYP2, NU107, ACOT8, S26A1, TB182, YTHD1, ASXL1, PI5PA, RIN3, MRTFB, RL37, KCNA2, RALA, STIM1, PITX1, IF4G2, SRPK2, RENBP, COG7, WNK1, SERF2, RPTN, SPSY, DAB2, RBM10, HNRPU, SPTB2, FOXK2, EWS, MEF2A, SP2, CO7A1, S30BP, NUCB1, ENL, IF4G1, K1C17, TLE3, TLE4, TOP1, SUH, CBG, ACK1, DEMA, AHNK, FOXO1, TROAP, BPTF, NFIA, ROA0, G3BP1, PABP4, ATM, PICAL, MAMD1, RIPK1, STIM1, MTMR1, CUL4B, ASPP2, KLF5, NFYC, CDK13, VEZF1, DSG2, TRI29, UBP2L, SRC8, PUM1, EPN4, RRP1B, NCOA6, DIP2A, MEF2D, NUMA1, R3HD1, KIF14, EBP, RCN1, KS6A1, RBMS2, TAF1C, NCOA2, SF01, JHD2C, MARE1, ELF2, TAB1, ZFHX3, ZYX, ADRM1, CCDC6, TAF9, STX1A, RFX7, QSER1, QRIC1, PRC2C, PBIP1, GSE1, TNR6A, CEAM5, Q3UKP4, COBL1, ARH40, SC31A, PEG3, SRBS2, Q3UU43, Q3UUE0, F91A1, ARBK2, Q497W2, Q4KL65, PHAR4, EPC2, CRTC2, BCORL, K2026, TGO1, PRC2B, TOIP1, SPG17, SHRM1, ZN362, LRIF1, RHG21, UBAP2, RBM26, RPRD2, ZN318, NCOR1, LAMA5, HCFC1, P3C2A, SAP, AP180, MAFK, SPTB2, SH3G1, ZYX, TSH3, INADL, WAPL, KAZRN, SBNO1, ARID2, DYH17, SAM9L, CDK13, LAR4B, BICRL, RHG21, HELZ, TTLL5, PANX2, PKHG2, NIPBL, LIN54, F135A, RPRD2, IF4G1, SPIC, SCYL2, NFRKB, INT1, ZN182, UGGG1, MDEAS, ZC3HE, RICTR, FIP1, CRTC3, SAS6, MCAF1, BCOR, GGYF2, NU188, CO039, UBN2, HAKAI, ASXL2, SPT6H, DDX46, KDM3B, PICAL, PRC2B, OOG2, ZIC5, NRK, POGZ, MAVS, CLAP1, EMSY, I2BP2, SRGP1, SH3R1, HUWE1, YTHD3, NU214, UBP2L, TMC5B, ZN598, TOPRS, SHAN2, Q80ZX0, ZNF18, Q810G1, BCL9L, LUZP1, PRSR1, DDX42, PALB2, P66A, GNS, LPP, TB10B, TGO1, Q8BIB6, ZN771, ZNT6, AAPK2, CNOT4, SP110, IFFO1, YTHD3, NCBP3, DEFI6, RBM14, CNOT2, CABS1, Q8C6L9, TCAL5, TAB1, SCYL2, ASPP2, PHC3, EPN2, PDLI5, I2BP1, RN135, AHNK2, NAV2, MISP, MGAP, ANKH1, PHAR4, XRN1, PELP1, Q8JZK6, Q8K0U8, AGFG1, TXD11, IL23R, ARHG6, SPART, SPICE, NUP93, CLASR, ZN786, SYNPO, FNBP4, ARFG1, ENAH, TNR6A, PHC3, SP20H, NAV1, VP37A, KMT2C, BD1L1, NUP35, STXB6, KNL1, TCAL3, MTSS1, SPART, NUP35, PUM2, STT3B, ALMS1, GEMI5, WIPF2, MAVS, UTP6, PI3R4, AMOT, P66B, STAG1, PCNP, LMO7, ATX2L, CSKI2, P66B, BBX, TITIN, HNMT, UBAP2, DCP1A, NRIF1, SMG7, RTF1, MAML1, ZN592, LAR4B, TAF4B, SHIP1, DDX17, RENT1, GPKOW, FUBP2, LPP, TTC28, PF21A, INT12, RCN3, CERS2, PDLI5, FUBP3, MY15B, ANCHR, CLP1L, Z512B, ZFR, EP400, NOL4L, RBM14, CIC, MED15, PIGS, DCR1C, SIN3A, MINT, EYA3, TEAD3, ATX2, RFC4, DHX58, ANX13, GORS2, TAB2, EPN4, ANR17, DPH2, WAC, DIDO1, YTHD1, AMRA1, TANC1, TXD12, F133B, RBM33, GPI8, Q9D2U0, ZC21B, FUND2, F162A, APMAP, Q9D809, FIP1, CNPY3, Q9DAV5, Q9DB24, ALG2, PLIN3, MYPT1, WWTR1, Q9EQC8, SALL1, RBP2, GILT, MFF, SP130, APC1, I2BPL, RBNS5, EPC1, ADNP, ZN106, TM245, CPVL, PTN23, WNK1, E41L1, ZHX3, ZN335, PKHG2, CCSE2, CQ10B, MLXIP, PKHA5, RC3H2, TAF9B, ZBT20, NCOA5, ZN532, APMAP, HYOU1, ADRM1, GIT2, BAG3, UBN1, PDLI7, DIAP3, RBM12, CARF, ETAA1, HXC10, TAB2, UGGG1, CDK12, ITSN2, CNOT2, TMEM9, DAPLE, NYAP2, KANL3, SON, LIMD1, KI21B, KI21A, PPIE, PCM1, GALK1, MRP5, SE1L1, LIMD1, TCF20, SUN2, AFF4, UBQL2, S30BP, NRBP, SIX4, TASOR, GMEB2, PARP4, NUP50, ZHX1, YETS2, HECD1, SCAF8, SRRM2, SCML2, S22AL, NCOR2, DEMA, POLH, R3HD2, ZN281, FBX7, RPGF2, IRS2, HYOU1, PRC2C, NCOR2, GMEB1, S23IP, SRPK3, Q9Z0I7, VNN1, KLK4, SE1L1, RGS6, E41L1
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Wang S, Yang F, Petyuk VA, Shukla AK, Monroe ME, Gritsenko MA, Rodland KD, Smith RD, Qian WJ, Gong CX, Liu T. Quantitative proteomics identifies altered O-GlcNAcylation of structural, synaptic and memory-associated proteins in Alzheimer's disease. The Journal of pathology 2017 243(1) 28657654
Abstract:
Protein modification by O-linked β-N-acetylglucosamine (O-GlcNAc) is emerging as an important factor in the pathogenesis of sporadic Alzheimer's disease (AD); however, detailed molecular characterization of this important protein post-translational modification at the proteome level has been highly challenging, owing to its low stoichiometry and labile nature. Herein, we report the most comprehensive, quantitative proteomics analysis for protein O-GlcNAcylation in postmortem human brain tissues with and without AD by the use of isobaric tandem mass tag labelling, chemoenzymatic photocleavage enrichment, and liquid chromatography coupled to mass spectrometry. A total of 1850 O-GlcNAc peptides covering 1094 O-GlcNAcylation sites were identified from 530 proteins in the human brain. One hundred and thirty-one O-GlcNAc peptides covering 81 proteins were altered in AD brains as compared with controls (q < 0.05). Moreover, alteration of O-GlcNAc peptide abundance could be attributed more to O-GlcNAcylation level than to protein level changes. The altered O-GlcNAcylated proteins belong to several structural and functional categories, including synaptic proteins, cytoskeleton proteins, and memory-associated proteins. These findings suggest that dysregulation of O-GlcNAcylation of multiple brain proteins may be involved in the development of sporadic AD. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
O-GlcNAc proteins:
SBNO1, F221A, CNOT1, WIPF3, MYO9A, APBB1, TAF4, P3C2A, DLGP1, C2C2L, DC1I1, CTRO, NCAN, ABLM1, KMT2D, MYPT1, PDZD2, RIMB2, TCPR2, ERC2, ANR28, LAMA5, OGT1, CLOCK, SI1L1, M3K13, HS12A, CP110, MTSS1, M3K7, TPD54, IF4G3, E41L2, FOXO3, KCNQ3, RNF13, MYPT2, AP180, PLIN3, MOT2, MAFK, HCN1, SLIT3, CLAP2, DEPD5, LIPA3, PP6R2, ANR17, LIPA4, NCOR1, GGYF1, HSBP1, ATRN, FLOT1, SYUG, TPPP, TOX4, SRBS2, TOX, ABLM3, AGFG2, RTN3, KCNH1, VAPB, ZFYV9, SC24B, CNOT4, ZMYM6, BAG3, DDAH2, ABL1, CRYAB, LMNA, MBP, FUCO, GCR, G3P, HSPB1, APOD, PERM, IF2A, NFL, NFM, SAP, VIME, CO4A, AN34C, ATX1L, ELFN1, DERPC, CALM1, CALM2, CALM3, LYAG, TAU, LAMC1, MTAP2, EPB41, BCR, LAMP1, PABP1, NFH, CO6A2, CO6A3, TPR, SKI, CSPG2, GFAP, KPYM, GNS, ZEP1, ARSB, ITB4, STMN1, ATF7, SYN1, NEUM, EGR1, SON, ELK1, TFE3, CSK22, ICAL, MAG, CSRP1, VDAC1, TENX, CBL, PTPRZ, OMGP, LAMA2, TENA, APC, DNJB2, MAP4, PTPRM, CLIP1, ZEP2, L1CAM, HS71L, CTNB1, ADDA, NU214, NUP62, SYUA, VATA, CUX1, STAT3, PTGDS, GRM5, GRIA1, MATR3, ATRX, NOTC1, MAP1B, SC6A8, PRC2A, UB2R1, NU153, RBP2, GSK3A, TAF6, TOB1, PPT1, FXR2, MECP2, HCFC1, AGFG1, NUP98, ATX1, DSRAD, LAMB2, CAD13, TPD52, STX17, MYPR, CSK21, F193A, SHPS1, IF4G2, PHC1, MAP1A, BASP1, RT36, FOXK1, PGBM, RHG04, HNRPU, SPTB2, ANK2, NAAA, DYST, NOTC2, TLE2, TLE4, NMDZ1, MEF2C, ZO1, ACK1, LG3BP, DEMA, CAMP2, AHNK, BPTF, CNTN1, ANK3, ROA0, GPS2, MTA1, LSAMP, GAB1, PICAL, ASAH1, RIPK1, KCC2B, BMPR2, VEZF1, UBP2L, GIT2, DPYL1, DCTN1, SRC8, PUM1, EPN4, RRP1B, NCOA6, KCNB1, LASP1, CRYM, NFIX, NUMA1, SHPRH, HECAM, IF4H, PLEC, PTPRR, PTPA, NCOA2, SF01, JHD2C, T22D1, PI5PA, TAB1, NCOA1, ZYX, SYUB, ADRM1, CCDC6, AINX, DPYL2, NTRK2, TBR1, RTN1, RFX7, QSER1, AAK1, QRIC1, MA7D1, TBC25, TB10B, TPRN, FIL1L, SVEP1, GRAP1, AMOT, CCD93, IGS11, ARMX4, NHLC3, PRC2B, SNP47, CE170, ZEP3, SKT, UBAP2, RBM26, AHDC1, VP13D, MRCKA, RPRD2, RN220, F1711, TASO2, MLIP, PAPD7, TNS2, KANK2, CRBG3, ANR40, ABLM2, CAPR2, LIN54, F117B, SCYL2, NFRKB, MDEAS, ZC3HE, LARP1, FIP1, PAMR1, MCAF1, MPRIP, GGYF2, NFXL1, PPR3F, K154L, RAPH1, UB2R2, HAKAI, MTSS2, ASXL2, CEP68, SPT6H, SYT13, MICA3, GRIN1, POGZ, ZFY16, MAVS, EMSY, RBBP6, SH3R1, HUWE1, YTHD3, FLIP1, LYRIC, RIMS1, F124A, LUZP1, PACS2, RLGPB, P66A, KCC1D, AHNK2, NAV3, TEX2, MGAP, CC28A, UBR1, ANKH1, NPAS3, MILK2, ULK2, PHAR4, DCP1B, SPART, CEND, RPTOR, KT222, ZN687, DOCK4, SYNPO, FNBP4, MINK1, OXR1, AVL9, CAMKV, SLAI1, NUP35, REPS2, NBEA, MYRIP, SVIP, PLBL2, NCOA7, TBC15, NEK9, DLG5, GEMI5, WIPF2, F222B, SMAP2, TM263, ZFN2B, LMO7, CTL1, ATX2L, PALLD, CSKI1, MADD, P66B, BAALC, ZCH14, LAR4B, PTPRU, RYR2, TAF4B, SYN2, DDX17, GPKOW, FUBP2, ARHG2, LPP, SNX21, MRTFA, SH3K1, PF21A, ATG2B, PLD4, REPS1, CYFP2, PHIPL, PGCB, PDLI5, MAP6, VCIP1, ZFR, EP400, SPT33, DOCK7, LRRC7, RBM14, NED4L, QKI, SYGP1, LMTK3, PLIN4, ARX, CIC, MED15, KKCC2, MINT, PKP4, ATX2, SH3G3, DPH2, KTNB1, TPPP3, SRBS1, BBS2, YTHD1, WNK3, CFA74, ZCHC2, TB182, AMRA1, ZC12C, SRCN1, SP130, BRD8, WDR13, EPC1, CA198, WNK1, E41L1, ZHX3, VIAAT, Z385D, DOCK5, GORS2, JUPI2, MA6D1, SIAE, PKHA5, RC3H2, TENS1, EMAL4, ZBT20, TANC2, E41LA, APMAP, ARFG3, GEPH, ENTP7, BMP2K, RN146, STAU2, T106B, CARF, TAB2, GGA3, ARHGC, SPN90, DLGP2, CAMP3, PHRF1, PLCE1, STOX2, KANL3, ARP21, S30BP, NRBP, CDC23, GGA2, AKA11, GMEB2, TNIK, PARP4, HCN2, MRTFB, YETS2, EPDR1, NOTC3, SPAT2, SYNRG, G3BP2, BSN, SCAF8, LIMC1, TRAK1, SHAN2, RIMS2, SRRM2, CTND2, JIP1, KCC2A, DLGP4, E41L3, GIT1, WNK2, C2CD2, TLN1, ARIP4, MTCL1, DCAF1, RPGF2, PRC2C, SHAN1, FLVC1, NCOR2, GMEB1, EMIL1, EPN1, NUMBL, M3K4, PCLO, ZHX2, S23IP
Species: Homo sapiens
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Hahne H, Sobotzki N, Nyberg T, Helm D, Borodkin VS, van Aalten DM, Agnew B, Kuster B. Proteome wide purification and identification of O-GlcNAc-modified proteins using click chemistry and mass spectrometry. Journal of proteome research 2013 12(2) 23301498
Abstract:
The post-translational modification of proteins with N-acetylglucosamine (O-GlcNAc) is involved in the regulation of a wide variety of cellular processes and associated with a number of chronic diseases. Despite its emerging biological significance, the systematic identification of O-GlcNAc proteins is still challenging. In the present study, we demonstrate a significantly improved O-GlcNAc protein enrichment procedure, which exploits metabolic labeling of cells by azide-modified GlcNAc and copper-mediated Click chemistry for purification of modified proteins on an alkyne-resin. On-resin proteolysis using trypsin followed by LC-MS/MS afforded the identification of around 1500 O-GlcNAc proteins from a single cell line. Subsequent elution of covalently resin bound O-GlcNAc peptides using selective β-elimination enabled the identification of 185 O-GlcNAc modification sites on 80 proteins. To demonstrate the practical utility of the developed approach, we studied the global effects of the O-GlcNAcase inhibitor GlcNAcstatin G on the level of O-GlcNAc modification of cellular proteins. About 200 proteins including several key players involved in the hexosamine signaling pathway showed significantly increased O-GlcNAcylation levels in response to the drug, which further strengthens the link of O-GlcNAc protein modification to cellular nutrient sensing and response.
O-GlcNAc proteins:
UBA6, SHOT1, MEX3A, TPC13, CNOT1, PGP, CE051, SMHD1, CC85C, I4E1B, ZBBX, B2R8K8, B2RB32, B3KNS4, B3KUR4, B4DF70, B4DIH0, B4DIR9, B4DTN6, B8XCX8, DX39A, BACH, GTPB1, SMAP, AP3B1, PSD12, PSMD9, ATOX1, PGRC1, SPT5H, TAF4, DFFA, CLIC1, EIF3F, WASL, IPO5, EF2K, PLOD2, MEIS1, PSDE, IMA4, BCL9, SDCB1, NOP56, IMA3, ARI1A, UBFD1, CCS, DVL2, KMT2D, ANM5, TNPO2, GEMI2, HAT1, MYPT1, GAK, XPO1, ZN609, SC16A, SR140, PUR4, NKRF, ZN185, CASC3, OGT1, PMM2, HMGB3, PPM1G, SHIP2, NVL, NUP42, R113A, KDM6A, DHX15, MCES, RRP8, SERA, DC1L2, ZW10, M3K7, RIPK2, WDR62, TXNL1, IF4G3, E41L2, FOXO3, DENR, XPOT, BUB3, ACTN4, HTSF1, SGTA, SYNC, LANC1, DHX16, ZNRD2, KPRB, AQR, GANP, HNRPQ, BUB1B, DIAP1, PLIN3, ANM3, CTND1, EIF1B, USO1, IF2P, NBN, SRGP2, N4BP1, ROCK2, CLAP2, CPNE3, BRE1B, ZC11A, CLU, T22D2, ANR17, GGCT, HMMR, VATG1, FLNB, NCOR1, SPAG7, PR40A, GGYF1, PSIP1, NDUS3, SRS10, KHDR3, SF3B1, CSDE1, PRKRA, KS6A5, NPM3, LYPA1, U520, TIPRL, OFD1, WDHD1, CD123, EIF3G, PSD10, SPF27, CIAO1, RMP, TOX4, SC24D, ST65G, UBXN7, PLPHP, NFAT5, SOGA1, UBP19, WDR47, SC31A, HEXI1, UBR5, SCAF4, UBE4B, ELP1, ZRAB2, KIF4A, VAPB, SNAPN, 6PGL, SMC2, ATG7, IPO7, AGM1, ZFYV9, SVIL, BAG4, AHSA1, PARN, PSMG1, SC24A, SC24B, AGRL2, TYDP2, PCNT, YETS4, STABP, ASML, EYA4, CEP43, HS74L, WIZ, STAU1, BAG2, BAG3, BPNT1, ECD, MBD3, BCL10, MOC2B, TOM40, CHK2, GSHR, PNPH, HPRT, ADA, CYTB, OAT, KITH, CPNS1, P53, HSPB1, TYSY, RPN1, RLA1, JUN, LA, NPM, NFL, NFM, ANXA2, SYEP, HS90A, HNRPC, SP1, 4F2, HS90B, ASNS, VIME, GSTP1, LEG1, HMGB1, UCHL1, LKHA4, H2A1, UBC, GLI2, ODP2, LAMP1, G6PD, ADHX, CDK4, SRF, PCNA, HARS1, ADT3, IMDH2, ACTN1, PEPD, XRCC5, LAMP2, RINI, EF2, PLST, ACPH, KAP2, CD99, GLU2B, AK1A1, KPYM, PO2F1, SYDC, ALDR, ERF3A, GNS, ZEP1, DESP, CREB1, GCFC2, UBF1, ATF7, CAN2, PYRG1, TCPA, SON, NELFE, ATF1, NFKB1, ICAL, IMDH1, GSTM3, FLNA, COMT, FBRL, PUR2, PUR6, UBA1, ENPP1, CBL, SP100, NFYA, SAHH, COF1, TENA, THTM, RS12, DNJB1, PSA4, DNMT1, RAE2, PTBP1, SYTC, 1433T, RFA1, APEX1, PYR1, MAP4, CALX, PSA5, NDUS1, TPP2, SHC1, TKT, PML, MARCS, PRDX6, PRDX5, PRDX3, KCY, 2AAA, 2AAB, CDC27, NMT1, SDHA, GDIA, METK2, DNJA1, AKT1, PUR9, HNRH1, S10AB, ARRB2, DCTD, TF2H1, KINH, CSTF2, DUT, TTK, PROF2, CAH8, RFC4, RFC2, HXD13, MYH9, MYH10, ADDA, BASI, NU214, DEK,