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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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Levine ZG, Fan C, Melicher MS, Orman M, Benjamin T, Walker S. O-GlcNAc Transferase Recognizes Protein Substrates Using an Asparagine Ladder in the Tetratricopeptide Repeat (TPR) Superhelix. Journal of the American Chemical Society 2018 140(10) 29485866
Abstract:
The essential mammalian enzyme O-GlcNAc Transferase (OGT) is uniquely responsible for transferring N-acetylglucosamine to over a thousand nuclear and cytoplasmic proteins, yet there is no known consensus sequence and it remains unclear how OGT recognizes its substrates. To address this question, we developed a protein microarray assay that chemoenzymatically labels de novo sites of glycosylation with biotin, allowing us to simultaneously assess OGT activity across >6000 human proteins. With this assay we examined the contribution to substrate selection of a conserved asparagine ladder within the lumen of OGT's superhelical tetratricopeptide repeat (TPR) domain. When five asparagines were mutated, OGT retained significant activity against short peptides, but showed limited limited glycosylation of protein substrates on the microarray. O-GlcNAcylation of protein substrates in cell extracts was also greatly attenuated. We conclude that OGT recognizes the majority of its substrates by binding them to the asparagine ladder in the TPR lumen proximal to the catalytic domain.
O-GlcNAc proteins:
E2F8, PRXD1, TSPY2, CGB1, ZN839, CENPX, PDLI1, WWP2, FOXN3, IMA4, DPYL4, ABLM1, MEIS2, HGS, GAK, PLS1, MAGB4, ARPC5, DC1L2, FGF16, WIPF1, FRS3, RNF13, ZN207, ENSA, SMCA5, CE104, OGA, AP180, PQBP1, E2F6, ADAP1, DNJC8, FLOT1, TPPP, SCEL, CELF2, RPGF3, CLD14, KLF8, BAG3, PAK4, IGHG3, MBP, NFIC, SNRPA, CEL3A, RU1C, IGLC1, MTAP2, ERG, DMD, IRF2, ENPL, FOSL2, F261, SDC1, ATF1, ATF3, ELK1, TFEB, RXRA, COMT, USF1, FGFR3, NR4A1, THAS, MARK3, ARNT, PSB4, RBMS1, FCERG, MPIP1, CSTF2, CD68, BMI1, COPB2, SOX5, RS19, ARL4A, ETV5, IPP2, STAT6, ABL2, KSYK, TFPI2, RFX5, RXRG, CDK8, GSK3B, TAF6, SPS1, DNLI3, DYN2, GALK1, AGFG1, SC24C, DLX1, HDAC4, CTBP2, KLF3, TIRAP, PPIA, CSK21, 3BP2, SSBP2, CDKL1, BORG5, TFAP4, ROR2, CACB1, CREM, MECOM, TLE4, FAK1, COEA1, MTG8, GABPA, GABP1, DERM, TLE5, DEMA, ASPH, PP1R8, LCP2, DUS4, MTAP, EI2BE, G3BP1, GRB10, MTA1, PPIL2, PICAL, DYR1A, FHL3, NCOA4, CAYP1, CIRBP, DGCR6, VGLL4, CAPR1, GRB7, CASL, MEF2D, LASP1, SPCS2, SEPT2, PCM1, SIX1, SF01, TRIP6, RHOH, TAB1, ADRM1, CCDC6, BATF, NFE2, MAR1, NRF1, SMTL2, LETM2, OD3L2, AX2R, AMOT, AKIR2, PRR30, LARGN, RGPS1, PRC2B, TEX30, SNP47, PHYD1, KANK4, EEIG1, CPTP, AHDC1, KHDR2, EF1DL, OLM2B, ANKS6, RHG17, CNDH2, GRHL2, HMBX1, JHY, PP4R4, JMJD6, SCYL2, GOLM2, RSBNL, PSRC1, DRIC1, KCD18, FIP1, SAS6, CLM9, CCBE1, DUSTY, E400N, TM1L2, UBP45, MARK2, KCTD9, MCM10, CC186, POGZ, FLIP1, TAF8, RM55, S1A7A, SYVN1, PHLB1, CASZ1, PIM3, GA2L3, DDX42, CEP57, SPAS2, Z280B, PDPN, KCC1D, TF2LX, LOXH1, ARI3B, APLF, CC28A, RHG12, SKA3, PHC2, HYCC2, CMIP, SMAP1, RUSD2, ABI1, SYCE1, RPTOR, BATF2, S35F6, KLH36, ANS4B, MARHA, ZSCA1, UXS1, NECP1, BBOF1, LIPB2, MISSL, PHC3, TMIE, NUP35, OR4D6, JOS2, WDR48, GABP2, DPTOR, ALAT2, BRSK1, GRP4, HELQ, BL1S5, BCAS4, LMX1A, SSH3, SH3R2, FBF1, DLRB2, HDAC7, STYX, GTSF1, NEUR4, CSRN3, AP2B, SHC3, RORB, STAM1, CELF1, ARHG1, CSN5, RBPMS, ZDH16, PMEPA, UTP4, FERM2, SOSB2, R51A1, CR025, LENG1, BIRC7, PBIR1, CC126, PF21B, NRBF2, EDC3, PDLI5, ANCHR, ZC3HA, PKNX2, EGLN2, MARK4, UB2E2, PRRC1, NOL4L, AP4AT, SCLT1, TOX2, QKI, PNMA5, YPEL2, NUDC1, RB40C, UBL7, CK2N2, OSBL9, PNKP, PLIN2, CDCA3, HCD2, SNAG, ARI3A, DPYL5, CG025, SOSB1, TCF25, COE4, MIC25, UTP23, ALKB7, RBM42, LRRC1, TRMO, MCRI2, RSRP1, BBLN, ASHWN, SARG, NADAP, SSBP3, PLVAP, OSB11, B2L13, STK31, DGC6L, RM01, KLC2, SIK2, REN3A, NELFA, PPDPF, ILRUN, ES8L2, KI3X1, SP14L, B2L12, PKHA3, PKHA1, TAF9B, ARNT2, SYTL2, CIP1, CASS4, HINT3, PRDM5, PAR11, MBNL1, BRWD1, KLC4, TB22B, MBNL3, NFIP2, ZSC32, TMOD3, UGGG2, CWC15, KANL3, RBPJL, DKK4, LIMD1, DPOLL, AFF4, UBQL2, PUF60, BAZ2B, GGA2, ING1, BORG2, AGO2, ZHX1, MUCEN, ANR50, PCDGK, POLI, LIMC1, SMAG1, BAIP2, RPC10, PPIL1, C2CD2, MTL5, TBL2, YTHD2, T10B, SNX14, RT18B, GMEB1, STON1, PCLO, ZHX2, S23IP
Species: Homo sapiens
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