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Jeon BC, Kim YJ, Park AK, Song MR, Na KM, Lee J, An D, Park Y, Hwang H, Kim TD, Lim J, Park SK. Dynamic O-GlcNAcylation governs long-range chromatin interactions in V(D)J recombination during early B-cell development. Cellular & molecular immunology 2025 22(1) 39627609
Abstract:
V(D)J recombination secures the production of functional immunoglobulin (Ig) genes and antibody diversity during the early stages of B-cell development through long-distance interactions mediated by cis-regulatory elements and trans-acting factors. O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins that regulates various protein functions, including DNA-binding affinity and protein-protein interactions. However, the effects of O-GlcNAcylation on proteins involved in V(D)J recombination remain largely unknown. To elucidate this relationship, we downregulated O-GlcNAcylation in a mouse model by administering an O-GlcNAc inhibitor or restricting the consumption of a regular diet. Interestingly, the inhibition of O-GlcNAcylation in mice severely impaired Ig heavy-chain (IgH) gene rearrangement. We identified several factors crucial for V(D)J recombination, including YY1, CTCF, SMC1, and SMC3, as direct targets of O-GlcNAc modification. Importantly, O-GlcNAcylation regulates the physical interaction between SMC1 and SMC3 and the DNA-binding patterns of YY1 at the IgH gene locus. Moreover, O-GlcNAc inhibition downregulated DDX5 protein expression, affecting the functional association of CTCF with its DNA-binding sites at the IgH locus. Our results showed that locus contraction and long-range interactions throughout the IgH locus are disrupted in a manner dependent on the cellular O-GlcNAc level. In this study, we established that V(D)J recombination relies on the O-GlcNAc status of stage-specific proteins during early B-cell development and identified O-GlcNAc-dependent mechanisms as new regulatory components for the development of a diverse antibody repertoire.
O-GlcNAc proteins:
MPEG1, CUL4B, DHX8, RHG27, VIR, PNISR, FRPD1, RENT2, LAS1L, ITIH4, THOC2, MMRN1, PYR1, YTDC2, PTPRB, ANR44, RBM25, OSBL8, DAAF5, SAFB1, KI67, DESP, YTDC1, UBE4A, NUMA1, MORC3, HXK2, LEG9, KNG1, UBE3A, DCTN1, DIAP1, U5S1, RL21, PHB2, CPSF2, DHX15, EXOC4, STAG2, AP1B1, PININ, HNRH1, SP100, GP1BA, ITB3, RL35A, SPT5H, DHX9, E41L2, BAZ1A, ZFR, ROA2, PRS6A, FA5, AFAM, COR1A, SP1, C1QR1, COX2, AMY1, CO3, CO4B, B2MG, HBA, HBB1, K1C10, K2C1, CFAB, ALDOA, TBA1B, TBA3, ITAM, K1C18, LDHA, LCK, APOA4, PTPRC, CFAH, TTHY, ANXA2, ALBU, A1AT1, SPA3K, HS90A, TRFL, ENPL, APOE, MDHM, GNAI2, RPB1, ITB1, PDIA1, NUCL, APOA2, PTPRQ, CALM2, EF1A1, 4F2, PARP1, PERM, FINC, HS90B, K2C8, ITA5, ITB2, TCPA, RL7A, GELS, ICAM1, DNMT1, S10A6, RL27A, RS16, RL7, RSSA, LMNB1, ANXA6, RLA0, CD44, LEUK, H12, CN37, AMPE, HS71L, G3P, LAMP2, HSP72, ENOA, PTBP1, PPIA, TPIS, LYZ1, PCNA, PTPRA, BASI, KS6A1, KS6A3, COF1, FAS, THRB, RL13A, BIP, VIME, PLMN, VTDB, A1AT2, CBL, AP1G1, EIF3A, EST1C, ITAL, CD11B, MCM3, RS2, CD19, UBF1, TLN1, EZRI, MOES, KLKB1, H2AX, VAV, NCKP1, MUG1, KIF2A, DPP4, PTN6, FETUA, C5AR1, CEAM1, CD68, ANT3, SYWC, KIF4, DPOLA, RAB5C, RAB18, CD22, TSP1, CALX, RFC1, PRDX1, RL12, RL18, DNLI1, GRP75, DYN2, RL28, MMP9, STAT1, STA5B, EPS15, TCPQ, MSH2, H14, H15, RAGP1, SIPA1, NSF, PRS7, BRCC3, NEDD4, CAPZB, RL6, RL5, RL13, RL36, KSYK, PERE, ROA1, MCM4, MCM5, SAHH, K2C6A, VATA, PA2G4, RAB7A, RL9, ADT2, IMA1, PON1, DPOD1, UBP10, KPYM, STAT6, RL10A, CEBPZ, PIPNB, MSH6, UBP5, ATPB, UBP25, NICA, ACTN4, EF2, OPA1, FOXP1, TPM2, WDHD1, ARPC4, RUVB1, PCBP1, ACTB, IF4A1, RS20, UB2D3, ARF3, RL26, RL27, RL37A, ARF4, HNRPK, RS7, PRS4, RS8, RS15A, RS14, RS23, RS18, RS11, RS13, SMD2, ARF6, PRS10, RS4X, RL18A, RL23A, RS6, H4, VATB2, RAB1A, RAN, RL23, RS24, RS25, RS26, RL30, RL31, RS3, RL8, PROF1, RL40, HSP7C, PHB1, RL22, RACK1, ACTS, TBA4A, TBB4B, 1433F, IMB1, M4K1, PKN1, STIM1, PYRG2, ROCK1, RAD50, PYRG1, TCPH, TCPB, TCPD, TCPE, TCPZ, TCPG, WNK1, RHOG, RL19, H33, BACH2, MCM2, MCM6, RS3A, ANX11, SMRC1, FUMH, ARVC, TBB5, APOA1, A1AT4, TYY1, HNRL2, LYAM3, TOP2A, APOH, TERA, UBA1, PLAK, ATPA, IKZF1, SPA3M, SMRCD, TOP1, RAC2, PYC, IF2P, CBG, ACADS, AMBP, PECA1, SSRP1, ZCH18, K2C80, PSA, PTCD3, NSUN2, EDEM3, MCM9, TMC5, HMHA1, HP1B3, GUAA, H2AV, SMCA4, PRC2C, MIDN, K1C26, K22E, PSMD1, BRE1B, ESYT1, AAK1, RHG17, EDC4, UBP19, GPD1L, ELNE, SC31A, IQGA2, K22O, ITB2L, C1TM, UN13A, PLCH1, PDS5B, CENPJ, DDX46, TR150, A16A1, EHMT1, MCTP2, RBM27, CYFP2, PSME4, MYO1G, LC7L3, PUR4, MYH1, LEO1, SIN3A, XRCC1, ODO1, HNRPD, SAMH1, HELLS, ARHG2, I17RA, PML, 2A5G, PPM1G, CFAI, CERU, CTCF, PRDX2, EZH2, HCFC1, PA1B3, ARHG1, PLSL, A2AP, HSP74, DSG1A, GSLG1, EWS, RAD21, FSCN1, GDIB, DDX5, HS105, ITIH2, ITA6, EI2BD, SERA, KINH, PDCD4, PZP, PRG2, MYH10, MCM7, NPM, PCBP2, CTR9, DDX3X, CD180, SPTB2, SPR1A, TIF1B, TFR1, RU17, SPT6H, NDUA4, IF4G2, MINT, RHG30, H2B1B, TOP2B, TPP2, AT2A3, H2A2C, VINC, PUR2, CLH1, SYMC, GNPTA, PDS5A, CDC5L, CE290, F120A, UBP7, JADE3, K1C42, K2C72, SR140, K2C73, S23IP, IF4G1, RBM26, P4R3A, U520, ABCF1, SMHD1, UGGG1, XPO1, ANO6, KIF15, KIF11, FHOD1, FKB15, PTN23, LPPRC, SMRC2, ECM29, CHD4, PK3C3, NUP98, GMIP, NFRKB, TEX2, UBE2O, KDM3B, CE162, CNOT1, CAND1, LARP1, VIP2, RS9, RL35, RS27L, 2AAA, SND1, ASAP2, IPO8, HUWE1, LC7L2, MBB1A, INT7, CTDP1, PP6R1, ELP1, DCAF1, CLAP1, SCRIB, PUM1, NU214, NAA15, FACD2, FBLL1, UBP2L, SYMPK, SIG10, DDX42, ANFY1, EFTU, TNPO1, ROA3, PLD4, SYAC, S2512, NU107, PTBP3, NRDC, ERC6L, GANAB, SP130, NUP93, SUN2, RCC2, IPO5, EMSY, ODP2, RBGPR, SYLC, SYQ, ECHA, RL24, CLAP2, CNDH2, PB1, FLNA, SYIC, IFIX, CIP2A, GEMI5, UBP47, CTL2, TBCD, POGZ, ANC2, KS6A5, EFL1, LCAP, DOCK8, CND2, IWS1, RBM14, DOCK2, UBA6, MIC60, UFL1, VCIP1, NUP88, NED4L, RPB2, AQR, SMC4, SMC2, SYEP, TCRG1, LONM, OGT1, CHERP, CCAR1, INT5, PYGB, COPA, PLCG2, INT4, EIF3B, BCLF1, K319L, URP2, DNM1L, NEK9, FCHO1, PAF1, IPO11, CND1, MATR3, PLCL2, DP13A, PO121, SF3A1, HNRPL, NU133, EIF3C, BST2, CD177, ADIPL, CDC16, STPAP, LRC8C, ACSF2, EVI2B, MYH9, UHRF1, VIGLN, ADPGK, PSMD2, HNRL1, AT1A1, MICA1, CCAR2, DX39A, SRSF4, K2C79, RFA1, HNRPU, S25A3, RBM39, SEC63, IPO4, SFPQ, ACLY, IF4A3, NDUS1, ATPG, DDX1, UBAP2, HEMO, IPO9, RBM5, PRP6, SMCA5, SP16H, TADBP, SF3B3, SYDC, PP6R3, C1TC, NOP2, PDE2A, KIF2C, K2C5, SIR1, XPO5, SMRD2, ECHB, ARP3, EMIL1, UN45A, ACON, DPP3, HSP7E, GTPB4, ARBK1, SRRT, SF3B1, NU155, RRBP1, DHX30, RL17, NUDC2, 6PGL, COTL1, RM18, TRAP1, AT5F1, RL14, XPOT, PRPS2, RRP44, SMC1A, SMUF1, SMC3, PUR9, SNX2, ROA0, RL11, GARS, RL15, MTREX, MMS19, HNRPM, SYRC, NH2L1, RL34, GRIFN, UB2V2, S10AE, CORO7, STAG1, CUL5, SC23B, CALL3, NOP56, RL4, EF1G, PRP4, QCR2, PELP1, AP2B1, XRN2, NVL, EIF3K, 6PGD, SYF1, EIF3F, XPO7, IPO7, RENT1, BCAP, PESC, ERAP1, VPS35, EHD4, TFP11, XPO2, PKHA2, RBP2, UBE4B, SHIP1, HRG, XPO4, AN32B, GTF2I, DYHC1, STK4, COPB, DDX21, ACINU, FLII, IQGA1, HYOU1, HIP1R, FMNL1, SACS, SART3, GIT2, MY18A, ITA2B, FAK2, CAF1A, K1C17, FETUB, PLEC, PO210, ADDA, PCLO, COPG1, UBQL2, H2AY, ZEB2, GALK1, SC11A, MTA2, PR40A, TIM, MYO1C, INSRR, MD1L1, PDC6I, PFKAP, CXA10, GANP, IF2G, ADNP, P5CS, SAE2, ARI1, DX39B, CLIC1, SYVC, AP3B1, ILF3, USO1, HNRPC, BAZ1B, K1C16, SNUT1
Species: Mus musculus
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Gonzalez-Rellan MJ, Parracho T, Heras V, Rodriguez A, Fondevila MF, Novoa E, Lima N, Varela-Rey M, Senra A, Chantada-Vazquez MDP, Ameneiro C, Bernardo G, Fernandez-Ramos D, Lopitz-Otsoa F, Bilbao J, Guallar D, Fidalgo M, Bravo S, Dieguez C, Martinez-Chantar ML, Millet O, Mato JM, Schwaninger M, Prevot V, Crespo J, Frühbeck G, Iruzubieta P, Nogueiras R. Hepatocyte-specific O-GlcNAc transferase downregulation ameliorates nonalcoholic steatohepatitis by improving mitochondrial function. Molecular metabolism 2023 75 37453647
Abstract:
O-GlcNAcylation is a post-translational modification that directly couples the processes of nutrient sensing, metabolism, and signal transduction, affecting protein function and localization, since the O-linked N-acetylglucosamine moiety comes directly from the metabolism of glucose, lipids, and amino acids. The addition and removal of O-GlcNAc of target proteins are mediated by two highly conserved enzymes: O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) and O-GlcNAcase (OGA), respectively. Deregulation of O-GlcNAcylation has been reported to be associated with various human diseases such as cancer, diabetes, and cardiovascular diseases. The contribution of deregulated O-GlcNAcylation to the progression and pathogenesis of NAFLD remains intriguing, and a better understanding of its roles in this pathophysiological context is required to uncover novel avenues for therapeutic intervention. By using a translational approach, our aim is to describe the role of OGT and O-GlcNAcylation in the pathogenesis of NAFLD.
O-GlcNAc proteins:
DJC25, TITIN, FIBA, AOXC, LEG9, CLCA, MTP, MYH11, NTCP, PRDX6, DLDH, HCD2, GLU2B, PRDX4, RL21, GSH0, HGD, AMACR, PHB2, DOPD, PSMD4, SRSF5, PAHX, AGT1, S27A2, BHMT1, ANXA3, AP1B1, CP4AE, TIM44, CALU, AL1A7, OST48, PGRC1, COPB2, NIPS1, RL35A, AT2A2, PSB5, DPM1, EIF3D, EF1B, NMT1, UGDH, DHB12, WDR1, RDH7, ROA2, COMT, JAM1, IDHC, CP8B1, COPE, RBM3, CP1A1, ADH1, COX1, COX2, CO3, HVM14, IGKC, IGHG1, B2MG, HA1B, HBA, HBB1, K1C10, NU3M, ATP8, K2C1, ALDOA, AATC, AATM, K1C18, LDHA, G6PI, MAOX, ANXA2, ALBU, SPA3K, HS90A, PDIA4, ENPL, APOE, SODC, MDHM, PDIA1, NUCL, PGK1, FRIH, SODM, EF1A1, CATB, THIO, GSTM1, RRAS, H10, GPX1, HS90B, MUP2, K2C8, CP2D9, OTC, TCPA, FABPL, PDCD6, RL7A, MYH8, GPDA, RL27A, RS16, RL7, MDHC, RSSA, CALR, GTR2, HSPB1, PSMD3, ANXA6, RLA0, GLNA, NDKA, CAH3, LEG3, SRP14, PH4H, MUTA, ASSY, G3P, ENOA, PTBP1, AP2A2, SBP1, COX7C, UDB17, PPIA, TPIS, PTPRA, CATD, COF1, FAS, GSTP1, RL13A, COX5B, COX41, BIP, PRDX3, VIME, CP2A5, TPM3, VTDB, TGM2, EIF3A, MOV10, CATA, PPIB, CP2DA, LKHA4, AL1A1, CO8A2, RS2, URIC, TLN1, MOES, RADI, 3BHS3, CTNA1, U2AF2, DHE3, SYSC, MA2A1, RL3, PDIA3, PSB8, ACOHC, APEX1, ADHX, MUG1, GRN, DPP4, PABP1, FRIL1, OAT, VTNC, GSTA3, CTND1, ACBP, SCP2, LA, DRG1, CP2F2, HYES, RAB18, FAAA, FBRL, CALX, PRDX1, RL12, RL18, HEM6, NCPR, HMGCL, GRP75, CAP1, TKT, INMT, RL28, ACSL1, ECI1, SEPT2, STAT1, PURA, PXMP2, TCPQ, H14, PLIN2, ACADM, MYO1B, STT3A, QOR, ALDH2, AL3A2, CAZA2, GSHR, RL6, RL29, RLA1, RL5, RL13, RL36, ANXA5, LMNA, CBR1, T23O, ROA1, HPPD, ODBA, DHB8, DHI1, SAHH, GLYC, K2C6A, VATA, ACADV, PA2G4, RAB7A, ACADL, RL9, DHB2, DHB4, GSHB, ADT2, THTR, PON1, KPYM, RIDA, CPT2, ST1A1, ST2A1, RL10A, ODB2, KPYR, MOT1, CNBP, IDHP, HMCS2, ABCD3, RAB8A, ADK, ATPK, CX6B1, CYB5, ATPB, CP2AC, FUS, EF1D, ACTN4, TM9S2, EF2, GGLO, IF5, ARPC4, EIF3E, PCBP1, ACTB, CDC42, IF4A1, RS20, UBE2N, ARP2, ARF3, ABCE1, RL26, RL27, RL37A, S61A1, ARF4, GABT, HNRPK, 1433G, RRAS2, RS7, PP1B, PRS8, RS8, RS15A, 1433E, RS14, RS23, RS18, RS29, RS11, RS13, RS4X, RL18A, RL23A, RS6, H4, RAB1A, RAN, RL23, RS15, RS24, RS25, RS26, RS28, RS30, GBB2, RL30, CYC, RL31, RS3, RL32, RL8, YBOX1, PROF1, RS27A, RAC1, HSP7C, TCTP, DNJA1, CH60, 1433Z, IF5A1, RS17, RS10, PHB1, RL22, RACK1, ACTS, 1433T, TBA4A, TBB4B, TBA1C, 1433F, IMB1, EBP, NFIX, PEBP1, HNRH2, ELAV1, NHRF1, NACAM, DHB5, TCPB, TCPD, TCPE, TCPZ, TCPG, RL36A, AP2M1, RL19, ISC2A, KHK, RS3A, PSME2, ANXA4, MYL11, RS5, GSH1, FMO3, CPT1A, FUMH, CATC, LYPA1, G3BP1, FMO5, TBB5, RLA2, PRDX5, RET4, A1AT4, HNRL2, SC23A, NDKB, TERA, UBA1, PLAK, ATPA, PPA5, CP2E1, PYC, ATP5I, CLUS, ANXA7, ACADS, LG3BP, HSDL2, ML12B, METK2, K22E, AGT2, RMD3, MCCB, SC31A, IQGA2, S27A5, DDX17, OPA3, GLSL, ACBD5, COPD, A1CF, ODO1, MYL6, HNRPD, PSB6, GCDH, IRGM1, SPB6, STIP1, CAPR1, VDAC2, VDAC3, VDAC1, COQ8A, PRDX2, ARGI1, PLSL, ABCD2, HCDH, GPAT1, GDIB, HPT, DDX5, HS105, LASP1, NPM, NNTM, PCBP2, DDX3X, SSRD, SPR1A, NDUA4, CYTB, VAT1, SBP2, EST3A, UD11, SPRE, BTF3, RGN, CH10, UD16, DHSO, CP2CT, GSTT1, GPDM, CLH1, F120A, IF4G1, ODBB, ABCF1, LPPRC, SRSF1, RS9, RS27, RL10, RL35, IF2A, RS27L, H2B1C, KAT3, 2AAA, ATPMK, MIC27, 3HAO, S22AI, SND1, OSTC, MTCH2, NDUAC, HNRPQ, TBB2A, LDHD, ACTN1, GVIN1, MOGS, FA98B, COEA1, FLNB, ACD11, BDH, EFTU, LPP, ROA3, ALAT2, AL8A1, S2512, ECHM, CHDH, IPO5, TM214, SRP68, ECHA, SYNC, RL24, ASPH, FLNA, DHPR, AOFB, SSDH, ACOT4, UD2A3, THIM, ERF1, CPSM, NAKD2, MIC60, PARP9, THIC, S2545, G6PE, SYEP, LONM, NT8F2, DPYD, AL4A1, PDLI5, COPA, CP2DQ, EIF3B, ACSL5, TXTP, UD3A2, GALM, COQ9, SDHA, HACD3, MATR3, NDUS8, ALAT1, THIL, EIF3L, HNRPL, EPIPL, SGPL1, AL1L1, BPHL, EIF3C, CMBL, QCR9, H2AJ, SDHL, HUTU, AK1CD, TKFC, GYS2, THIKB, CGL, DHB13, ABHEB, EST1D, EST3B, ACSF2, MYH9, VIGLN, PSMD2, AT1A1, HNRPU, S25A3, SEC63, OASL1, SFPQ, TRAM1, CK054, ACLY, ACSM1, NDUS1, RINI, RMXL1, ATPG, DDX1, MGST1, CBR4, GCSP, NDUS2, EIF3H, KMO, SYYC, EST1F, BAAT, GCKR, DCXR, METK1, NADC, FTCD, GLYAT, TM205, ALDOB, ATLA3, ARLY, RPN1, NDUV1, GRHPR, PCCA, UGPA, FPPS, FADS1, TADBP, ETFD, THIKA, TRFE, PDIA5, SYDC, C1TC, MARC2, LRC59, PDIA6, ATAD3, SFXN2, ROAA, S14L2, THTM, STML2, SFXN1, GORS2, ECHB, ARP3, NONO, PLST, AASS, GLO2, ACON, DPP3, DHRS1, 3HIDH, IF2B, DHRS4, SARDH, NDUAA, ETFA, RTCB, PARK7, NDUS5, DNJA3, PCCB, MCCA, PECR, NUDT7, RTN4, AAAD, RRBP1, GDIR1, NDUA5, ATP5L, RL17, AMPL, DECR, MTAP, QCR8, NDUA2, SDHB, NDUB4, SAR1B, PCYOX, NDUB5, NDUB9, TXD17, TRAP1, AT5F1, ACO13, PPP6, SC61B, MTNA, RER1, 1433B, CYB5B, MAGT1, NDUA6, RL14, M2OM, TMM33, UCRI, VKOR1, OCAD1, ARPC2, MARC1, PUR9, DDAH1, ROA0, KYNU, CENPV, RL11, TECR, SERB1, QCR1, C560, RL15, GLYM, AL1B1, CISY, RS19, ODPB, HNRPM, LMAN1, PGM1, SYRC, CY1, SYTC, GAL3A, CNDP2, TMEDA, FKB11, ERP44, HYEP, HACD2, ATPD, PLCC, NDUV2, GHC1, IDH3A, MSRA, GLGB, ACAD8, PRPS1, SAPC2, IPYR, RL37, SOX, U2AF1, RL4, EF1G, ATPO, IAH1, QCR2, HUTI, CSAD, AL7A1, CP27A, AP2B1, RPN2, PGAM1, COASY, ECHP, KCY, M2GD, NDUA9, NDUS7, RT11, 6PGD, SSRG, EIF3F, NDUA8, PUR6, GSTK1, NB5R3, ASPDH, MTL26, NDUBA, NDUS3, HOGA1, RMD1, ETFB, ATP5H, KEG1, TMT1B, LACTB, RENT1, DHB11, MMSA, DPYS, ERAP1, VPS35, MVP, SET, RT29, PYGL, IVD, TMOD3, NIT2, COPB, AK1A1, CMLO1, PLS1, RL38, PXMP4, GNA1, AL9A1, CD2AP, STA10, TRXR1, RHOA, PSA6, K1C17, F16P1, HACL1, GNMT, S2513, CLIC4, VKGC, DIC, EIF3I, COPG1, TBL2, ACOX1, ESTD, DEST, TEBP, MPU1, SQOR, PSA1, TPSN, RUVB2, KAD3, HAOX1, PDC6I, PROD, SYFB, SUCA, PREB, ECI2, ENTP5, DECR2, TAGL2, ORNT1, PACN2, EHD1, MAAI, IF2G, BPNT1, AIFM1, DX39B, SYVC, ILF3, STRAP, HNRPC, PX11A, LETM1, SUCB2, SUCB1, PSA7, HNRPF, MCAT
Species: Mus musculus, Mus spicilegus
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Narayanan B, Zahra F, Reeves RA, Aggarwal A, O'Meally RN, Henry RK, Craven M, Jacobson A, Cole RN, Kohr MJ, Umapathi P, Zachara NE. Differential Detection of O-GlcNAcylated proteins in the heart using antibodies. Analytical biochemistry 2023 678 37507081
Abstract:
Thousands of mammalian intracellular proteins are dynamically modified by O-linked β-N-acetylglucosamine (O-GlcNAc). Global changes in O-GlcNAcylation have been associated with the development of cardiomyopathy, heart failure, hypertension, and neurodegenerative disease. Levels of O-GlcNAc in cells and tissues can be detected using numerous approaches; however, immunoblotting using GlcNAc-specific antibodies and lectins is commonplace. The goal of this study was to optimize the detection of O-GlcNAc in heart lysates by immunoblotting. Using a combination of tissue fractionation, immunoblotting, and galactosyltransferase labeling, as well as hearts from wild-type and O-GlcNAc transferase transgenic mice, we demonstrate that contractile proteins in the heart are differentially detected by two commercially available antibodies (CTD110.6 and RL2). As CTD110.6 displays poor reactivity toward contractile proteins, and as these proteins represent a major fraction of the heart proteome, a better assessment of cardiac O-GlcNAcylation is obtained in total tissue lysates with RL2. The data presented highlight tissue lysis approaches that should aid the assessment of the cardiac O-GlcNAcylation by immunoblotting.
O-GlcNAc proteins:
A0A023T778, A0A075B5P3, A0A075B5P4, A0A075B5P6, A0A075B5T2, A0A075B5T7, A0A087WP81, A0A087WPL5, A0A087WR50, A0A087WS16, A0A087WS46, A0A087WSC6, A0A0A0MQ79, A0A0A0MQA5, A0A0A0MQD2, A0A0A0MQF6, A0A0A0MQJ4, A0A0A0MQM0, A0A0A6YVU8, A0A0A6YW67, A0A0A6YW80, A0A0A6YWP6, A0A0A6YX26, A0A0A6YXF6, A0A0A6YXH3, A0A0A6YXV1, A0A0B4J1E7, A0A0B4J1H7, A0A0F6AIX5, A0A0G2JDI9, A0A0G2JDL9, A0A0G2JDR9, GBP6, A0A0G2JDW2, A0A0G2JE25, A0A0G2JEP0, A0A0G2JEX1, A0A0G2JEY6, A0A0G2JF52, A0A0G2JF85, A0A0G2JG74, A0A0G2JGJ1, A0A0G2JGS0, A0A0J9YU07, A0A0J9YU24, A0A0J9YUZ4, A0A0M3HEQ0, A0A0N4SV40, A0A0N4SV66, A0A0N4SVB1, A0A0N4SVB8, A0A0N4SVK8, A0A0N4SVL9, A0A0N4SW07, A0A0R4J023, A0A0R4J038, A0A0R4J050, A0A0R4J052, A0A0R4J093, A0A0R4J0H8, A0A0R4J0I1, A0A0R4J0K2, A0A0R4J0L5, A0A0R4J0L6, A0A0R4J0S3, A0A0R4J0T5, A0A0R4J0X5, A0A0R4J0Z1, A0A0R4J138, A0A0R4J196, A0A0R4J1E2, A0A0R4J1J3, A0A0R4J1Q6, A0A0R4J1R7, A0A0R4J1W0, A0A0R4J1W7, A0A0R4J260, A0A0R4J275, A0A0U1RPW2, A0A0U1RPY8, A0A140LHG8, A0A140LIU4, A0A140LIZ5, A0A140LJ98, A0A140LJB7, A0A171EBL2, A0A1B0GQU8, A0A1B0GR11, A0A1B0GRP7, A0A1B0GRV0, A0A1B0GRV3, A0A1B0GRW3, A0A1B0GS58, A0A1B0GS68, A0A1B0GS70, A0A1B0GS91, A0A1B0GSG5, A0A1B0GSK8, A0A1B0GSR9, A0A1B0GX27, A0A1C7CYV0, A0A1C7ZN10, A0A1D5RM79, A0A1L1SQ51, A0A1L1SQA8, A0A1L1SS27, A0A1L1SST0, A0A1L1STE6, A0A1L1SV25, A0A1L1SV73, A0A1W2P6E6, A0A1W2P6P1, A0A1W2P768, A0A1W2P7A1, A0A1W2P812, A0A1Y7VJY8, A0A1Y7VKT9, A0A1Y7VKY1, A0A1Y7VLP0, A0A286YCI8, A0A286YCS6, A0A286YD12, A0A286YDA2, A0A2I3BPW0, A0A2I3BQE3, A0A2I3BQF4, A0A2I3BRL8, A0A2K6EDJ7, A0A2R8VHF9, A0A2R8VHX0, A0A338P6D9, A0A338P6G0, A0A338P6N4, A0A338P703, A0A338P731, A0A338P786, A0A338P7D7, A0A338P7F1, A0A338P7G4, A0A3B2W824, A0A3B2WB97, A0A3B2WBC6, A0A3B2WCN9, A0A3Q4EBV4, A0A3Q4EG54, A0A3Q4EH93, A0A3Q4EI56, A0A494B908, A0A494B923, A0A494B955, A0A494B985, A0A494B9D8, A0A494B9P3, A0A494B9W3, A0A494B9X1, A0A494B9Y4, A0A494BA51, A0A494BAJ6, A0A494BB38, A0A494BBC1, A0A494BBG8, A0A571BEC9, A0A571BEG4, A0A571BG59, A0A571BGD8, A0A5F8MPB9, A0A5F8MPD6, A0A5F8MPK9, A0A5F8MPM4, A0A5F8MPN8, A0A5F8MPW1, A0A5F8MPY2, A0A5K1VVQ9, A0A668KL51, A0A668KLD3, A0A6I8MWZ8, A0A6I8MX27, A1BN54, A2A513, A2A5F5, A2A6F8, A2A6Q8, A2A7A7, A2A813, A2A848, TM201, A2A997, A2A9Q2, OBSCN, A2AAW9, A2AE89, A2AEX6, A2AFQ0, A2AFQ2, A2AGJ9, A2AI69, A2AI87, NDUF6, A2AIM4, A2AIW9, A2AK42, A2AKD7, A2AKU9, A2AKV9, A2AL12, A2ALV7, CAVN4, A2AMW0, UBR4, A2AN84, A2APD7, A2AQN4, MYH7B, A2AQR0, A2AQY8, A2AS98, TITIN, A2AT02, A2AU61, A2AUD5, RBGP1, A2BDW0, A2BH06, A2BI12, A2CEK3, A2CES4, A2CG35, A3KFU5, A3KGU5, ARMT1, A6X925, A8DUK4, A8JYK8, CISD3, B1AR28, B1AR93, B1ARA3, B1ASE2, B1ASZ3, B1AT10, B1AU25, B1AUX2, B1AV14, B1AXW5, B1AZ14, B1B0C7, B2C3G8, GNAI1, B2RXT3, B2RY24, B7FAU9, B7ZCI2, B8JJI4, B8JJM5, B8JK32, B9EIZ7, D3YTL5, D3YU39, D3YUG3, D3YUK5, D3YUM1, D3YUP1, D3YVF4, D3YVI6, D3YVV9, D3YW19, D3YWA4, D3YX99, D3YXT0, D3YY36, D3YYE1, D3YYG9, D3YYS6, D3YZ06, D3YZ71, D3Z041, D3Z067, D3Z0G0, D3Z0X5, D3Z0Y2, D3Z1B5, D3Z1V4, D3Z263, D3Z2B3, D3Z2Y8, D3Z4A4, D3Z5K6, D3Z5W3, D3Z627, D3Z636, D3Z6F5, D3Z780, D3Z7A7, D3Z7D5, D3Z7K3, D3Z7U0, D3Z7V3, D5MCW4, D6REH1, D6RFU2, D6RFU9, D6RGM3, D6RH37, E0CXB1, E0CXH2, E0CXN7, E0CXS3, E0CY23, E0CZ90, E0CZE0, E9PUD2, E9PUE8, E9PUM3, E9PUU2, E9PUY9, E9PV12, FIBA, E9PV63, E9PV66, E9PVP0, E9PVU0, E9PWE8, E9PX89, E9PYF1, E9PYI8, E9PZ00, E9PZ88, E9PZC4, E9PZD8, E9PZF0, E9PZI9, E9PZP8, E9Q133, E9Q1J7, E9Q1T9, E9Q1V0, E9Q264, RYR2, E9Q4T8, ACACB, DESP, E9Q5B5, E9Q5I9, E9Q616, E9Q6A9, E9Q6C2, E9Q6W2, E9Q7A5, E9Q800, E9Q933, E9Q9C0, E9Q9T8, E9QJV4, E9QKY4, E9QL13, E9QN70, E9QPD7, E9QPX3, F6QPR1, F6QYE1, F6QYF8, F6RR81, F6VVE6, F6VVY4, F6VY18, F6XI62, F6YCA7, F6ZV59, F7A3N3, F7DBQ0, SKI3, F8VPN4, F8VQJ3, F8WGL3, F8WHU8, F8WIB1, F8WIE5, F8WIT2, F8WIV2, F8WIV5, F8WJ05, G3UVU2, G3UVV4, G3UW30, G3UW85, G3UWN9, G3UX26, G3UXI6, G3UXX3, G3UYR8, G3UZ33, G3UZ48, G3X8R0, G3X8R1, G3X8T3, G3X8T9, QNG1, G3X956, G3X975, G3X977, G3X983, G3X9Q1, G3X9U9, G3XA48, G5DDB7, G5E823, G5E839, G5E883, G5E8R7, H3BIY9, H3BJI5, H3BJP9, H3BJQ7, H3BKH4, H3BKH6, H3BL49, H3BLH2, H7BWZ3, H7BX01, H7BX88, H7BX99, I7HLV2, J3QMG3, J3QPW1, J3QPZ9, J3QQ13, K3W4R2, K3W4S6, K4DI63, L7N451, M0QWU8, CAN2, DPYL2, DLDH, GSTO1, RL21, AMACR, SCRB2, PSMD4, ECH1, ANXA3, C1QBP, TIM44, AL1A7, CAVN1, SYUA, IMPCT, DSG2, NIPS2, AT2A2, PGAM2, PDLI1, AOC3, FHL2, PSA3, DHB12, WDR1, MTX2, ROA2, COMT, BIRC6, IDHC, GNPI1, KBL, AFAM, C1QR1, LANC1, ADH1, COX1, CAH2, CO3, CO4B, HBE, NU1M, NU4M, NU5M, FABP4, MYG, ALDOA, KAPCA, AATC, AATM, TBA1B, CO5, G6PI, MAOX, TTHY, KCRM, ANXA2, ALBU, K2C4, HS90A, CBR2, ENPL, MDHM, PDIA1, NUCL, PGK1, FRIH, SODM, H2AZ, IFI5B, ANXA1, EF1A1, NID1, CATB, THIO, RRAS, H2B1M, H10, LIPL, FABPH, HS90B, DMD, K2C8, ITPR1, TCPA, UCP1, PFKAL, RL7A, GELS, CAH1, GPDA, AT1B1, RS16, RSSA, CALR, HA2B, PSMD3, LMNB1, GLNA, PMGE, CAH3, LEG1, LEG3, CN37, DDX3L, AMPE, ENOA, SBP1, TPIS, CATD, FAS, SERPH, COX41, BIP, PRDX3, VIME, PLMN, ENOB, VTDB, TGM2, AP1G1, EIF3A, CXA1, CRYAB, EST1C, CATA, PPIB, GSTA4, LKHA4, AL1A1, RS2, LYN, TLN1, EZRI, MOES, CTNA1, PTMA, DHE3, PSMD7, FKB1A, PDIA3, ACOHC, ADHX, PGS1, PGS2, NP1L1, ATX10, MUG1, OAT, FKBP4, HMGB2, DESM, MP2K1, SCP2, LA, ANT3, SYWC, MIF, RAB21, HSPB7, TSP1, ODPA, CALX, RL12, PPM1B, TAGL, HMGCL, GRP75, CAP1, TKT, VP26A, INMT, ECI1, H14, ALDR, ALD2, COF2, ACADM, NSF, VPS4B, PRS7, RB11B, NEDD4, PA24A, ALDH2, GSHR, MK14, PFKAM, RL6, ANXA5, ABCD1, LMNA, CBR1, COX8B, TNNI3, ADT1, HEP2, ROA1, INPP, MCM4, CAV1, SAHH, FMO1, GDIA, CSRP3, VATA, ACADL, DHB4, MLRV, HDGF, ADT2, THTR, PON1, KPYM, NDUS6, CPT2, ODB2, IDHP, RD23A, PUR8, DDX6, ADK, ACYP2, AP4A, CX6B1, UBP5, ATPB, CD38, ERP29, EF2, TPM1, B2L13, RUVB1, EIF3E, PCBP1, NPL4, ACTB, CDC42, IF4A1, UBC12, UBE2N, ARP2, ARF3, RL27, 1433G, RRAS2, RS7, PP1B, RS8, RS15A, 1433E, RS23, SMD3, EF1A2, RS4X, AP2S1, RS6, H4, RAN, RS24, RS26, CYC, RL32, FBX40, PROF1, LIS1, HSP7C, TCTP, CH60, IF4E, 1433Z, RS17, PHB1, ACTC, RACK1, 1433T, TBB4B, 1433F, USP9X, IDHG1, NACAM, F16P2, DCUP, PYRG1, TCPB, TCPE, TCPZ, ARF5, AP2M1, RL19, ISC2A, CSRP1, NRP1, RS3A, CPT1A, WBP2, FUMH, LAMA4, TBB5, G6PD1, APOA1, A1AT5, APOH, TERA, UBA1, PLAK, MYH6, KCRB, CO6A1, FABP5, ATP5I, CBG, MPRI, ACADS, AMBP, CD36, Q14BI5, ACSS3, A1BG, TRI72, Q20BD0, NLR1A, HSDL2, OSBP1, CUL4A, Q3TG45, Q3TGM7, GUAA, TM38A, Q3TXN1, PSMD1, SC31B, SYRM, DDB1, Q3U2G2, Q3U422, Q3U816, Q3U8Y1, Q3UER8, Q3UF75, TM10C, Q3UGX2, HAP28, MYLK3, Q3UJQ9, GPD1L, PP1R7, HDGR2, SC31A, THEM4, Q3UZJ4, ST1D1, Q3V117, HSPB6, D39U1, Q5SQ27, Q5SQB0, Q5SUS9, Q5SWN2, ACACA, MYH4, MYH1, COPD, A1AG1, ODO1, LAMA2, STIP1, VDAC1, COQ8A, ADIPO, 2A5G, LAMA5, HARS1, HCDH, GDIB, HS105, SERA, PZP, MIME, POSTN, PON3, DAG1, MYOM1, SPTB2, TX261, TIF1B, TSN, TFR1, NDUA4, CYTB, CAVN2, BTF3, CH10, ADH7, CP4B1, TOP2B, TPP2, H2A2B, H2A2C, NQO1, VINC, PUR2, CTNA3, CLH1, RFTN1, NOMO1, K2C73, 2ABA, U520, XPO1, KCRS, Q6P8N8, OXSR1, LPPRC, UBR2, CAND1, CAND2, MLEC, RS9, IF2A, KAT3, 2AAA, ATPMK, Q7TNL5, SRCA, ELP1, SESD1, FLNB, PANK4, IREB2, EFTU, MIRO1, SAM50, S2540, SYAC, SYNM, F210A, S2512, F13A, EHD2, ECHM, SYIM, AHSA1, RCC2, IPO5, ODPX, MAON, ODP2, ECHA, PPA6, Q8BRB6, AAPK2, Q8BTS3, CSN7B, PPME1, NUDT9, AOFB, SSDH, THIM, KANK2, VP13C, COMD2, PGM5, NHLC2, GPCP1, ASGL1, LCAP, EXOG, THOP1, Q8C483, EMC1, Q8C845, Q8C8G3, ANK2, VWA8, ARK72, SYEP, LONM, UN45B, BICRL, PGP, FBX4, AL4A1, PDLI5, DI3L2, PYGB, FERM2, COPA, PAK2, HMCS1, ACAD9, MIRO2, AFG32, EIF3B, ACSL5, FIBB, TACO1, PDXK, COQ9, SDHA, FMO2, LZIC, HIBCH, THIL, ERF3A, SUOX, FAHD1, S2542, EIF3C, CMBL, COQ6, Q8R2P8, FABD, GBRL1, CLYBL, TMC2, UBP15, RMC1, MAVS, EST1D, ACSF2, CK068, SYLM, MYH9, PPCS, HNRL1, AT1A1, LMCD1, HNRPU, S25A3, RBM39, FLNC, ETFR1, Q91VB8, NDUS1, HNMT, DDX1, CBR4, EIF3H, CISD1, HEMO, VPS36, RHG35, FAKD4, L2HDH, RPN1, TWF1, MYH7, PCCA, UGPA, SMCA5, HNRLL, ETFD, TRFE, RHEB, MACD1, SYDC, MPI, RT02, CPT1B, DCMC, SMRD2, EIF3M, ECHB, Q99JZ4, RRAGC, VMA5A, ACON, DCTN2, NAMPT, PPIF, DHRS1, 3HIDH, DHRS4, MAT2B, NDUAA, ETFA, DDAH2, RTCB, GRPE1, NDUS5, DNJA3, ASPN, MCCA, PPR3A, RM09, ACS2L, NUDT7, RTN4, GDIR1, PRP8, NDUA5, COX6C, ATP5L, LGUL, AMPL, MYL9, NDUC2, DECR, MTAP, PKP2, NDUA2, SDHB, NDUB4, TM223, COTL1, NDUB9, TXD17, TRAP1, AT5F1, ACO13, 1433B, DPH2, M2OM, CHSP1, NAR4, TPPP3, MIC19, SMC1A, NUD17, PUR9, SNX2, CYGB, ROA0, COQ5, MITOS, MPPB, CYBP, BIEA, QCR1, HS12B, RL15, GLYM, EFTS, AL1B1, CISY, 5NT3A, ODPB, SYRC, EXOS7, CY1, SYTC, HINT2, ILEUA, GAL3A, HHATL, RM53, ODO2, OXSM, CUL2, EFHD1, PLCC, NDUV2, FUND2, F162A, ECHD3, IPYR, QCR7, ITPA, RL4, EF1G, T126A, PHP14, MTU1, ATPO, QCR2, PLIN3, LMAN2, PGAM1, ACDSB, OCTC, RT15, 6PGD, NDUA8, PUR6, GSTK1, MECR, NDUBA, NDUS3, CRIP2, RMD1, ETFB, LACTB, DHB11, MMSA, ERAP1, VPS35, RM46, EHD4, PARVB, M3K20, PSMG2, PYGL, IVD, DYHC1, NQO2, ACTN2, COPB, AK1A1, DDX21, RL38, SH3L1, PROF2, TBA8, PRELP, IQGA1, HYOU1, LDB3, GLTP, PALS2, PPCE, MLRA, DRG2, PLEC, S2513, TOM40, CLIC4, DNJA2, ACOT2, AAKB1, DEST, ACOT9, KAD1, PSA4, PSB3, PSB2, SAE1, BMP10, MYO1C, KAD2, KAD3, CUL1, PDC6I, COR1C, SUCA, KAD4, CATZ, ABEC2, CBPQ, EHD1, S12A7, GRK1, MTNB, GBP2, SYUG, TWF2, NFS1, NDUA7, CLIC1, STK39, MTMR9, SUCB2, SUCB1, PCKGC, PSD10, MCAT, S4R1F2, S4R1N1, S4R242
Species: Mus musculus
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Zhu WZ, Palazzo T, Zhou M, Ledee D, Olson HM, Paša-Tolić L, Olson AK. First comprehensive identification of cardiac proteins with putative increased O-GlcNAc levels during pressure overload hypertrophy. PloS one 2022 17(10) 36288343
Abstract:
Protein posttranslational modifications (PTMs) by O-GlcNAc globally rise during pressure-overload hypertrophy (POH). However, a major knowledge gap exists on the specific proteins undergoing changes in O-GlcNAc levels during POH primarily because this PTM is low abundance and easily lost during standard mass spectrometry (MS) conditions used for protein identification. Methodologies have emerged to enrich samples for O-GlcNAcylated proteins prior to MS analysis. Accordingly, our goal was to identify the specific proteins undergoing changes in O-GlcNAc levels during POH. We used C57/Bl6 mice subjected to Sham or transverse aortic constriction (TAC) to create POH. From the hearts, we labelled the O-GlcNAc moiety with tetramethylrhodamine azide (TAMRA) before sample enrichment by TAMRA immunoprecipitation (IP). We used LC-MS/MS to identify and quantify the captured putative O-GlcNAcylated proteins. We identified a total of 700 putative O-GlcNAcylated proteins in Sham and POH. Two hundred thirty-three of these proteins had significantly increased enrichment in POH over Sham suggesting higher O-GlcNAc levels whereas no proteins were significantly decreased by POH. We examined two MS identified metabolic enzymes, CPT1B and the PDH complex, to validate by immunoprecipitation. We corroborated increased O-GlcNAc levels during POH for CPT1B and the PDH complex. Enzyme activity assays suggests higher O-GlcNAcylation increases CPT1 activity and decreases PDH activity during POH. In summary, we generated the first comprehensive list of proteins with putative changes in O-GlcNAc levels during POH. Our results demonstrate the large number of potential proteins and cellular processes affected by O-GlcNAc and serve as a guide for testing specific O-GlcNAc-regulated mechanisms during POH.
O-GlcNAc proteins:
MA7D1, CAVN4, OTUD4, FIBA, TRDN, DPYL2, CLCA, MYH11, KNG1, PRDX6, AKAP1, DLDH, NDUBB, GSTO1, CASQ2, RL21, PHB2, ECH1, NDUA1, TIM44, CAVN1, AKAP2, SLK, NIPS2, AT2A2, PGAM2, EF1B, ATX2, NMT1, XIRP1, PDLI1, MYPC3, SNX3, DC1I2, PLIN4, ROA2, RAD, CLPP, TOM1, COX1, COX2, CAH2, CO3, IGJ, KV2A7, IGKC, GCAB, IGHG1, IGH1M, B2MG, HBA, HBB1, LAMC1, FABP4, CFAB, MYG, ALDOA, ANF, AATC, AATM, TBA1B, LDHA, G6PI, TRY2, TTHY, KCRM, ANXA2, ALBU, SPA3K, ENPL, APOE, MDHM, ITB1, PDIA1, NUCL, PGK1, FRIH, MYL3, SODM, NDUB1, ANXA1, EF1A1, CATB, TAU, THIO, GSTM1, H2B1F, H10, CO1A1, FABPH, HS90B, DMD, PFKAL, COX5A, RL7A, GELS, MYH3, AT1B1, GLUT4, RL7, MDHC, RSSA, CALR, HSPB1, ANXA6, GLNA, B4GT1, GSTM2, H12, LDHB, SPTN1, G3P, ENOA, HXK1, PPIA, TPIS, BASI, COF1, RL13A, SERPH, COX5B, COX41, BIP, PRDX3, VIME, CYTC, ENOB, TGM2, EIF3A, CBX3, CXA1, PIMT, CRYAB, CATA, CAPG, GSTA4, RS2, TLN1, MOES, RADI, CTNA1, DHE3, FKB1A, MAP4, RL3, H2AX, PDIA3, PABP1, FRIL1, FETUA, DESM, AIMP1, LA, ANT3, RANG, MIF, PTN11, HSPB7, ODPA, CALX, PRDX1, RL12, RL18, FBLN2, HMGCL, GRP75, CAP1, TKT, RL28, ACSL1, ECI1, H14, H11, H15, H13, ALDR, COF2, ACADM, PRS7, ADX, ALDH2, CAPZB, RL6, RL29, CACP, RL13, ANXA5, TBCA, LMNA, CX7A2, TNNI3, ADT1, ROA1, PCY1A, CAV1, ODBA, CSRP3, ACADV, PA2G4, TNNT2, ICAL, ACADL, CAV3, MLRV, ADT2, LUM, KPYM, NDUS6, CPT2, RL10A, ODB2, CCHL, MOT1, IDHP, STOM, ADK, ATPK, ACYP2, ATP68, ATP5E, AT5G2, CX6B1, CX7A1, COX7B, CYB5, UBP5, ATPB, WFS1, ACTN4, EF2, OPA1, TPM1, B2L13, PCBP1, ACTB, RS20, PPLA, UB2D3, UBC12, UBE2N, RL26, RL27, SUMO2, 1433G, RS7, RS8, 1433E, RS14, RS18, RS11, RS13, DLRB1, EF1A2, RS4X, RL23A, RS6, H4, RAN, RS15, RS25, RS30, RL30, CYC, RL31, RS3, RL32, RL8, FBX40, YBOX1, RS27A, HSP7C, MPC1, CH60, GNAS2, 1433Z, HMGB1, IF5A1, ACTG, ACTH, RS12, RS10, RL22, ACTC, UB2L3, TBA4A, TBB4B, H31, IMB1, PEBP1, HINT1, IDHG1, NACAM, TCPD, SGCD, SGCA, WNK1, RL19, SRSF3, H32, RS3A, G3BP2, ANXA4, COQ7, G3BP1, LAMA4, QCR6, PRDX5, APOA1, CO1A2, NDKB, TERA, UBA1, MYH6, ATPA, KCRB, CO6A1, PGBM, EMAL1, ATP5I, CLUS, ANXA7, ACADS, CD36, NEBL, PERM1, TRI72, HSDL2, HP1B3, PRC2C, TM38A, Q3TV00, SRSF6, FUBP2, SDHF1, EI3JA, LIMC1, AAK1, NDUB6, MCCB, COBL1, SLMAP, SRBS2, K22O, CPZIP, NDUF2, MYPN, HSPB6, MLIP, IASPP, TM1L2, ODO1, LAMA2, STIP1, REEP5, VDAC2, VDAC1, COQ8A, LAP2B, PRDX2, HCFC1, LAMB2, HSP74, HCDH, FBN1, FXR1, KTN1, GDIB, DDX5, KINH, LASP1, PZP, NPM, NNTM, SNRPA, SPTB2, SPEG, SRBS1, DBNL, NDUA4, FKBP3, IF4G2, ZYX, CAVN2, SPRE, SF01, CD34, CH10, H2A2B, H2A2C, NQO1, VINC, EI3JB, CLH1, H2A2A, GPSM1, IF4G1, KCRS, LPPRC, AT1A2, CAND2, RS9, CMYA5, FHOD3, ATPMK, MIC27, MSRB2, NP1L4, MTCH1, MTCH2, PICAL, NDUAC, HNRPQ, HUWE1, LC7L2, MIC10, NEXN, SRCA, LNP, CLAP1, SRA1, UBP2L, NRAP, BDH, GLRX5, ATPF1, EFTU, H2A3, LPP, MYPT2, IF4B, ECHM, RCN3, SYIM, EIF2A, ODPX, EEA1, ODP2, ECHA, COQ3, RL24, FLNA, TIDC1, PLIN5, SYP2L, SSDH, THIM, MIC60, PABP2, BOLA3, SYEP, LONM, H2A1F, H2A1H, H2A1K, SEPT8, PGP, AL4A1, SLAI2, PDLI5, PYGB, PAK2, AFG32, EIF3B, FIBB, COXM2, COQ9, SDHA, SIR5, ACD10, NDUS8, NNRE, HIBCH, THIL, MARE2, QCR9, H2AJ, DC1L1, SPART, NAR3, MIC13, CLYBL, PP14C, TXLNB, MAVS, MYH9, VIGLN, PSMD2, AT1A1, LMCD1, HNRPU, S25A3, FLNC, SFPQ, NDUS1, MIC25, ATPG, SH3L3, UBAP2, NDUS2, EIF3H, CISD1, HEMO, EGLN1, L2HDH, RPN1, NDUV1, GRHPR, MYH7, PCCA, UGPA, ETFD, THIKA, TRFE, TOIP1, MACD1, CLIP1, K2C5, UBXN1, ALPK3, RT02, CPT1B, TALDO, ROAA, THTM, STML2, PACN3, ECHB, PLST, ACON, DCTN2, NAMPT, PPIF, NDUAA, ETFA, GRPE1, PARK7, NDUS5, DNJA3, PCCB, MCCA, PPR3A, EH1L1, ACS2L, RRBP1, GDIR1, NDUA5, COX6C, TOM22, ATP5L, NDUB2, COXM1, RM24, NDUC2, DECR, QCR8, NDUA2, FIS1, SDHB, NDUB4, NDUB5, NDUB9, AT5F1, RS21, ACO13, 1433B, CYB5B, KGD4, NDUA6, NDUB3, PSMD9, RL14, NDUB7, M2OM, UCRI, MIC19, OCAD1, PIN4, NDUS4, RT28, SERB1, SPCS2, SSBP, QCR1, NSF1C, C560, CISY, TOM70, RS19, ODPB, HNRPM, PGM1, SCOT1, CY1, HINT2, GAL3A, MCEE, CHCH2, ERP44, NOL3, MMAB, ODO2, COA3, RT33, ATPD, NDUB8, NDUV2, IDH3A, F162A, ARMC1, RL37, QCR7, RL4, EF1G, EFHD2, PRS37, ATPO, QCR2, PGAM1, MYPT1, LNEBL, TELO2, NDUA9, NDUS7, NDUA8, NDUBA, NDUS3, CRIP2, ETFB, ATP5H, MIC26, MMSA, EHD4, NDUAD, POPD1, HRG, PALLD, JPH2, IVD, NHRF2, PALMD, ACTN2, AK1A1, DBLOH, MYOZ2, PDK2, HSPB8, HIG1A, BAG3, AUHM, MACF1, VAPB, NDRG2, ACOT2, QKI, PRS30, UBQL2, H2AY, GLYG, ACOX1, DEST, KAD1, PSA1, KAD2, KAD3, CAD13, PYGM, IF4H, COR1B, SUCA, ECI2, SH3BG, TAGL2, PACN2, EHD1, AIFM1, NDUA7, BAG6, USO1, PLM, LETM1, SUCB2, SUCB1, K2C6B
Species: Mus musculus
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Wu JL, Chiang MF, Hsu PH, Tsai DY, Hung KH, Wang YH, Angata T, Lin KI. O-GlcNAcylation is required for B cell homeostasis and antibody responses. Nature communications 2017 8(1) 29187734
Abstract:
O-linked N-acetylglucosamine (O-GlcNAc) transferase (Ogt) catalyzes O-GlcNAc modification. O-GlcNAcylation is increased after cross-linking of the B-cell receptor (BCR), but the physiological function of this reaction is unknown. Here we show that lack of Ogt in B-cell development not only causes severe defects in the activation of BCR signaling, but also perturbs B-cell homeostasis by enhancing apoptosis of mature B cells, partly as a result of impaired response to B-cell activating factor. O-GlcNAcylation of Lyn at serine 19 is crucial for efficient Lyn activation and Syk interaction in BCR-mediated B-cell activation and expansion. Ogt deficiency in germinal center (GC) B cells also results in enhanced apoptosis of GC B cells and memory B cells in an immune response, consequently causing a reduction of antibody levels. Together, these results demonstrate that B cells rely on O-GlcNAcylation to maintain homeostasis, transduce BCR-mediated activation signals and activate humoral immunity.
O-GlcNAc proteins:
FAIM3, BLTP1, BCORL, M3K15, KANL3, EXC6B, PLHD1, CTTB2, MYO1E, SCLT1, TAF4B, TCOF, FLOT1, OXLA, HDAC1, SYPL1, SEM4D, MA2B1, PPE2, PLD3, DPOD2, NOCT, HNRH1, API5, DFFA, MMP8, DPM1, EIF3D, ESS2, CTNL1, VTI1B, S28A2, FA5, CO4B, IGKC, LAC1, IGHA, IGHDM, HA11, LAMC1, TBA1B, LDHA, HVM51, SPTA1, ZFP1, EGR1, ENPL, RPB1, ITB1, ENV1, 4F2, HS90B, HA2B, HB2A, CD44, BLK, CN37, LAMP2, ZFP37, PTBP1, HB2I, BASI, FAS, EVI2A, MDR1A, BGAL, ITAL, LYN, TLN1, MOES, U2AF2, MAP4, GNA13, RL3, CATG, DPP4, PTN6, HEXA, NKTR, HMGB2, SUH, CEAM1, GTR3, DRG1, RAB5C, CD22, FMR1, VGFR1, GRP75, CAP1, ECI1, FOXK1, STAT1, NKX25, TCPQ, H11, H13, IL12B, CAPZB, RL5, VDR, RET3, ADCY7, VA0D1, AAAT, IMA1, STOM, FUS, NICA, RU2A, EF2, AAAS, RUVB1, ABCE1, DCAF7, 1433G, ACTA, RS6, VATB2, RL23, RL8, PP2BA, RACK1, TBB4B, M4K1, ITPR3, SURF6, ELAV1, EVL, H2B1A, AT8A1, TCPH, TCPB, NXN, TBB5, HNRL2, CREB1, PLAK, 3MG, CO6A1, LG3BP, COE1, CNN2, NSUN2, HMHA1, SNUT2, SMCA4, TPC10, TGRM2, I20L2, LMF1, PUF60, ZSWM8, PRRC1, SC31A, CPZIP, ITAD, ULK4, ITA1, DYHC2, LIN54, JKIP3, GRHL3, MYO1G, SIN3A, IRAG2, SAMH1, KHDR1, LY75, RASA3, NPT2A, CAPR1, ARHG2, PML, IMA5, LAP2B, PRP4B, M4K2, TS101, PLSL, CTNA2, VSX2, CD37, SERA, PCBP2, TIF1B, COCH, NUP62, RALY, UT14A, ARG39, CLH1, ATS16, F120A, NOP58, TEDC2, U520, RRP12, SMHD1, ANO6, TTBK1, CHD4, SARM1, NUP98, RASL2, TNKS1, AT1A2, NFRKB, DDX55, DNA2, H2B1C, CMYA5, GIMA8, CYFP1, SPAG5, HNRPQ, RPF1, MBB1A, PRC2A, ADCY2, MOGS, SDA1, FA98B, WIPI2, TRRAP, XYLT1, WDR82, GNS, ERLN2, S38A9, WASF2, S2512, NIM1, TBL1R, ZN526, CARF, HES7, UNC80, RBGPR, ECHA, ELMO1, ATOSB, KMT2C, FLNA, TPC2, RBBP5, POGZ, DOC10, SYFA, SMKZ, COR2A, RBM14, DOCK2, CASP9, RAE1L, NUP88, RPB2, UACA, SYEP, P66A, VPS50, COPA, VWF, TXTP, ZN536, LMBD1, R4RL1, C2D1A, URP2, STX5, GT251, SDHA, PO121, ABLM1, COL12, ALAT1, RORB, PDLI2, ERO1B, CD177, PSPC1, NUP58, STAB2, LRC8C, COX18, MAVS, PLBL1, UN93B, EVI2B, MYH9, ESIP1, VIGLN, PSMD2, HNRL1, CCAR2, SP7, RECQ5, SFXN3, IF4A3, RINI, DDX1, UBAP2, S15A4, DNJC9, MASP2, UXS1, CSCL1, BMP2K, CYRIB, SYDC, C1TC, GLYR1, PDIA6, CIC, S12A6, ATAD3, MYO5A, MCLN1, ABEC3, STML2, SFXN1, PRP19, TARA, MCRS1, RTCB, NDUS5, S12A9, SF3B1, ANR17, NU155, TR34A, BAP1, PRP8, NUDC2, TSN31, RN138, RTRAF, RU2B, YETS4, M2OM, MIC19, SNX2, DDX28, CXXC1, RUSD4, ILF2, CHTOP, LUC7L, DIM1, MCES, SEC13, SP2, NOP56, U2AF1, EF1G, MCEM1, EVPL, PRP4, CMTR1, WWP2, DHB11, PESC, TLR9, IRX6, KRT81, RBP2, AFF4, KAT2B, STK3, NUP50, DDX21, ACINU, SIGIR, ZN207, SLAF1, SON, H2AY, MTA2, SAE1, MYO1C, RUVB2, TRPV2, PFKAP, ARC1B, ASAH1, VAPA, EHD1, IF2G, CLIC1, HNRPF
Species: Mus musculus
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Gurel Z, Zaro BW, Pratt MR, Sheibani N. Identification of O-GlcNAc modification targets in mouse retinal pericytes: implication of p53 in pathogenesis of diabetic retinopathy. PloS one 2014 9(5) 24788674
Abstract:
Hyperglycemia is the primary cause of the majority of diabetes complications, including diabetic retinopathy (DR). Hyperglycemic conditions have a detrimental effect on many tissues and cell types, especially the retinal vascular cells including early loss of pericytes (PC). However, the mechanisms behind this selective sensitivity of retinal PC to hyperglycemia are undefined. The O-linked β-N-acetylglucosamine (O-GlcNAc) modification is elevated under hyperglycemic condition, and thus, may present an important molecular modification impacting the hyperglycemia-driven complications of diabetes. We have recently demonstrated that the level of O-GlcNAc modification in response to high glucose is variable in various retinal vascular cells. Retinal PC responded with the highest increase in O-GlcNAc modification compared to retinal endothelial cells and astrocytes. Here we show that these differences translated into functional changes, with an increase in apoptosis of retinal PC, not just under high glucose but also under treatment with O-GlcNAc modification inducers, PUGNAc and Thiamet-G. To gain insight into the molecular mechanisms involved, we have used click-It chemistry and LC-MS analysis and identified 431 target proteins of O-GlcNAc modification in retinal PC using an alkynyl-modified GlcNAc analog (GlcNAlk). Among the O-GlcNAc target proteins identified here 115 of them were not previously reported to be target of O-GlcNAc modification. We have identified at least 34 of these proteins with important roles in various aspects of cell death processes. Our results indicated that increased O-GlcNAc modification of p53 was associated with an increase in its protein levels in retinal PC. Together our results suggest that post-translational O-GlcNAc modification of p53 and its increased levels may contribute to selective early loss of PC during diabetes. Thus, modulation of O-GlcNAc modification may provide a novel treatment strategy to prevent the initiation and progression of DR.
O-GlcNAc proteins:
UBP36, PYR1, RBM25, TRIPC, CAN2, PRDX4, AIP, GSTO1, X3CL1, DHX15, HNRH1, AT2A2, GPX4, TRI18, O88568, HA1W, LDHA, ANXA2, RIR1, HS90A, ANXA1, HS90B, TCPA, UMPS, H12, LEG1, DDX3L, TPIS, HB2I, FAS, IMDH2, MCM3, PSB8, SYWC, PRDX1, DNLI1, CAP1, STAT3, PURA, TCPQ, H14, H11, NEDD4, CAPZB, RL36, HS74L, ADT1, MCM4, RAB7A, ADT2, KPYM, RL10A, ACTN4, EF2, RIPK2, SF3B6, PCBP1, ABCE1, 1433E, RS11, 1433Z, TBA1A, EM55, HNRH2, ELAV1, TP53B, TCPB, ARF1, PSME2, TBB5, UBA1, CNN2, TGM3, INF2, GUAA, FUBP2, LRRF1, DDX17, SRC8, MYL6, SAMH1, KHDR1, IRGM1, SPB6, VDAC3, ASNS, LAP2A, PPM1G, PSN2, ZN638, FXR1, SERA, PZP, NPM, GPDM, NOP58, TPM4, NEST, TNKS1, RS9, MBB1A, ATX2L, FLNB, SYAC, LSS, CKAP4, HEAT3, THOP1, AROS, TXNL1, SYEP, STAU2, AGO4, HMCS1, PNPT1, ANLN, MATR3, WIPI1, MYH9, PARN, AT1A1, FLNC, RAB14, SFXN3, IPYR2, SMC1B, TADBP, LRC59, NONO, GRPE1, NU155, OGFR, MTAP, CYB5B, MPPB, GARS, HNRPM, TRIR, IPYR, EF1G, PUR6, RAI14, LIMA1, PALLD, TMOD3, DDX21, WDR12, CUL3, TRXR1, DREB, ZBP1, TOM40, DNJC7, IIGP1, DEST, ACOT9, KAD2, TEN1, VAPA, GBP2, P5CS, SAE2, SYVC, HNRPF
Species: Mus musculus
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Trinidad JC, Barkan DT, Gulledge BF, Thalhammer A, Sali A, Schoepfer R, Burlingame AL. Global identification and characterization of both O-GlcNAcylation and phosphorylation at the murine synapse. Molecular & cellular proteomics : MCP 2012 11(8) 22645316
Abstract:
O-linked N-acetylglucosamine (O-GlcNAc) is a dynamic, reversible monosaccharide modifier of serine and threonine residues on intracellular protein domains. Crosstalk between O-GlcNAcylation and phosphorylation has been hypothesized. Here, we identified over 1750 and 16,500 sites of O-GlcNAcylation and phosphorylation from murine synaptosomes, respectively. In total, 135 (7%) of all O-GlcNAcylation sites were also found to be sites of phosphorylation. Although many proteins were extensively phosphorylated and minimally O-GlcNAcylated, proteins found to be extensively O-GlcNAcylated were almost always phosphorylated to a similar or greater extent, indicating the O-GlcNAcylation system is specifically targeting a subset of the proteome that is also phosphorylated. Both PTMs usually occur on disordered regions of protein structure, within which, the location of O-GlcNAcylation and phosphorylation is virtually random with respect to each other, suggesting that negative crosstalk at the structural level is not a common phenomenon. As a class, protein kinases are found to be more extensively O-GlcNAcylated than proteins in general, indicating the potential for crosstalk of phosphorylation with O-GlcNAcylation via regulation of enzymatic activity.
O-GlcNAc proteins:
A0JNY3, A2A653, A2A654, TANC2, ZEP3, MA7D2, CKAP5, CAMP1, LZTS3, A2AJ19, AJM1, MA7D1, A2ALK6, RPGP1, UBR4, A2AP92, SKT, ANR63, A2ATK9, A2AUD5, A2BI30, A6H6J9, A6MDD2, A8DUV1, B1AQX6, B1AR09, GRIK3, B1ATI9, B1AWT3, NHSL2, FRS1L, UBP24, DLGP4, B2RQ57, B2RQ80, PYR1, B2RQL0, B2RQQ5, GNAI1, B2RUE8, OTU7B, B2RWX1, B6ZHC4, B6ZHC5, B7ZCA7, B7ZMP8, B7ZNA4, B7ZNF6, B7ZWM6, B9EHE8, CTTB2, B9EKL9, PTPRZ, D1FNM8, D3YU59, D3YWX2, DGKH, D3YXR8, PGBD5, SHAN1, D3Z0V7, D3Z2J5, D9HP81, E0CYT1, E9PUA3, E9PUC4, DGKD, E9PUR0, E9PV14, E9PV26, KI67, E9PWL1, E9PWM3, E9PY55, E9PZP8, E9Q1M1, E9Q2B2, E9Q3D6, E9Q3G8, E9Q3M9, E9Q4N6, E9Q616, E9Q6T8, E9Q6Y8, NUMA1, E9Q828, E9Q9I2, E9Q9J6, E9QA16, E9QAP7, E9QAR5, SC16A, E9QJU8, E9QMJ1, SYGP1, RFIP2, HXK2, CAN2, SC22B, DPYL2, STXB1, TCOF, DCTN1, GLU2B, EF2K, PRDX4, AIP, NUMBL, GSTO1, GSH0, M3K5, PSMD4, DHX15, NPC1, BMPR2, VIAAT, BCAT2, CTND2, PITM1, CSK22, REPS1, ACK1, SLK, CAC1B, PGRC1, IMPA1, SYUA, AKA7A, STRN, RL35A, AT2A2, PGAM2, ATX2, NMT1, E41L2, GPX4, EMC8, DHB12, HCN4, KDM6A, ZN326, SORL, GRPE2, KLC1, ZFR, O88568, HCN2, HCN1, BSN, TOM1, RPP30, DNJB5, COX1, HA1D, HBA, K2C1, MBP, ALDOA, PGFRB, LDHA, G6PI, ENPP1, NEUM, ANXA2, RIR1, HS90A, EGR1, MDHM, KCC4, NFL, NFM, GNAI2, PDIA1, NUCL, CADH1, RC3H2, LRC4B, IGS11, DERPC, UBB, IFI5B, IFI4, ANXA1, EF1A1, H2B1F, PARP1, HS90B, DMD, KCC2A, TCPA, A4, COX5A, GELS, UMPS, NCAM1, GPDA, MDHC, SRP54, RLA0, GLNA, H12, LEG1, DDX3L, SPTN1, AP2A2, TPIS, KS6A3, COF1, GNAO, NFH, SERPH, VIME, MTAP2, EIF3A, CBX3, IMDH2, MCM3, CTNA1, MAP4, GNA12, GNA13, PDIA3, PSB8, NCKP1, PABP1, FKBP4, HMGB2, AIMP1, LA, ACM4, SYWC, RANG, RAB5C, RAB18, CALX, PRDX1, RL12, PPM1B, DNLI1, CAP1, STAT3, PURA, OPRM, TCPQ, CX6A1, MSH2, H14, H11, ALDR, ALD2, CBP, AINX, NEDD4, RP3A, CAPZB, SRPRB, RL36, SOX2, HS74L, ADT1, ROA1, INPP, PCY1A, MCM4, CSRP3, RAB7A, CDN2A, HDGF, ADT2, IMA1, UBP10, KPYM, RIDA, HMGA2, RL10A, CCHL, SOX1, RAB2A, ATX1, CACB3, HMCS2, GOGA3, ATPK, ATPB, ACTN4, IDI1, ACOT8, PTPA, KCNN2, KCNN3, TB10A, TB182, SF3B6, MRTFB, DOCK4, MYPR, EIF3E, PCBP1, LIPA3, ACTB, IF4A1, SNP25, RAB10, CSN2, RRAS2, PRS8, RS15A, 1433E, RS18, RS11, SMD1, ABI2, EF1A2, ACTA, VATB2, RL23, RS24, GBB1, HSP7C, TCTP, GNAS2, 1433Z, HMGB1, IF5A1, ACTG, RS17, RS12, UB2L3, RACK1, ACTS, TBA4A, TBA1A, TBB4B, PLXA2, DCC, EBP, NFIX, EM55, HNRH2, NCOA1, ELAV1, RGRF2, USP9X, TCPB, TCPE, TCPZ, NUCB2, IRS2, WNK1, RL36A, CSRP1, SEPR, RS3A, DPYL1, MPRIP, CAC1A, ATP5J, BOP1, RS5, WBP2, CXAR, PLPL9, G3BP1, RBBP6, CDS1, TBB5, IL6RB, NMDE2, NMDE3, TOP2A, NOTC1, NDKB, AQP1, UBA1, CTNB1, S30BP, NFIA, NUCB1, MARK3, APLP1, ENAH, ATPA, TF65, YES, MARK2, PGBM, PYC, CAPR2, EMAL1, LARP7, BAX, CNN2, LYAR, CHD8, CNNM1, INF2, TT21B, Q0IJ77, TRIO, VGF, TANC1, CDK12, Q14B66, MA6D1, NSUN2, MCM9, PHAR1, PSD3, Q2Q7P0, FILA2, Q3TAD4, NB5R4, GUAA, METK2, PRC2C, Q3TRG3, PLPL6, K22E, YETS2, Q3TY93, FUBP2, F117B, Q3U882, LBR, TM109, FOXK2, Q3UFK1, Q3UGZ4, TNR6C, DAB2P, ZEP2, AAK1, Q3UHT7, DTX3L, EDC4, PARP3, WASC4, GRIN1, Q3UQ23, SRBS2, THSD4, MRCKA, SPRY3, KSR2, GRM5, TBCD9, LRRF1, ARMX5, STOX2, SHAN3, UBN1, OXR1, DDX17, PHAR4, ANR28, ZN608, Q571B7, PRAG1, TAB3, Q58DZ3, IQEC2, Q5DU62, AAPK1, NUFP2, UNKL, SMG7, RBM27, CYFP2, TM1L2, PSME4, ANR40, Q5SUH6, GGNB2, SYNRG, Q5SVJ0, RPGP2, TBC9B, ACACA, Q5SXC4, Q5XJV5, LMTK3, RN123, ZDHC8, SRC8, MYL6, SKI, SAMH1, IRGM1, CLD11, NPT2A, SPB6, VDAC2, VDAC3, VDAC1, STYX, RBBP4, ASNS, NCOA2, LAP2A, PPM1G, ASTN1, PRDX2, HCFC1, APC, KCNA4, AP180, FXR1, GDIB, GRID2, GRID1, CBX5, SERA, LASP1, NPM, PCBP2, M3K7, SRBS1, DBNL, SH3G1, CYTB, IF4G2, MINT, ZYX, RALY, TFE3, Q640L6, AR13B, HECAM, NPDC1, SYN2, TBR1, ISG15, ABCG1, ATP4A, MRC2, G3PT, PTN13, TPP2, CTNA3, SBNO1, BEGIN, K1549, GIT1, SLAI1, PKP4, PEAK1, CDK13, SH3R1, MYOF, ABLM3, ARMX2, CE170, LAR4B, NOP58, Q6GR78, TPM4, NIPBL, RRP5, FBX41, Q6NVA3, RPRD2, WWC2, ZN532, Q6NXW0, S23IP, SMHD1, NEST, CSKI1, Q6P9N8, MTSS2, AHDC1, PTN23, TRAK1, SRSF1, CHD4, DLGP3, NUP98, NYAP1, KCC2D, AT1A3, AT1A2, NFRKB, RIGI, MAGI1, WDFY3, TACC1, GGYF2, PF21A, KDM3B, CNOT1, LARP1, Q6ZQB7, NU188, Q6ZQJ9, Q6ZQK4, RS9, RL10, IF2A, SC6A5, SEM6D, 2AAA, EEIG1, MTCH2, PICAL, MRO2B, SCN4B, PLPR4, HNRPQ, TBB2A, SMAP2, Q7TNS5, PLPR3, MBB1A, LNP, TPPP, ATX2L, OTUB1, EXOS3, MAP6, ELP1, SI1L2, LRRC7, ERBIN, PHF24, R3HD2, NAV3, AGRL3, Q80TS6, AUXI, MADD, AVL9, PUM1, UBP8, NU214, SEPT9, NAA15, CAMP3, FA98B, TDRKH, EPN1, TMCC2, AGFG2, UBP2L, Q80X68, C2C2L, FLNB, LRRT4, WNK3, PRIC2, CNKR2, ZN598, SHAN2, AGRB3, Q80ZX0, ZFYV1, MAST4, RHG32, ENTP3, LPP, PEF1, ACTBL, TET3, MYPT2, IF4B, SYAC, F168A, TBL1R, TB10B, CSTP1, CARF, TGO1, FRM4A, SYIM, ANS1B, DLGP2, ZNT6, RCC2, ABLM2, LSS, UNC80, NOE2, CF015, EMSY, ODP2, GGA3, SYLC, DMXL2, IMP2L, CLAP2, LIPA2, ASPH, CNOT4, FLNA, F163B, GEPH, CREST, KCC1D, PGES2, KANK2, GEMI5, IFFO1, OSBL6, YTHD3, TM266, POGZ, LACC1, MAP1S, A16L1, SI1L1, PP4R4, MYO9A, THOP1, RBM14, Q8C2R1, CNOT2, Q8C6E9, CC134, ANK2, ELFN1, DIDO1, NHSL1, WDR37, DCTN4, SYNPO, BCAS3, VCIP1, Q8CE98, TAB1, SCYL2, NED4L, SYEP, F193A, GNAL, OGT1, NAV1, SYNJ1, RPGF2, EP400, PHC3, P66A, TBCE, VWF, STAU2, LIN7A, TBC23, ZBT20, RTN1, HS12A, DNM1L, UNC5B, UNC5A, ANLN, AGFG1, MATR3, Q8K314, AHI1, NDUS8, I2BPL, PREP, ABLM1, EIF3L, ERF3A, HNRPL, IQEC1, DOCK7, DC1L1, SPART, BST2, RFIP5, AT2A1, NUP35, LUZP1, MAVS, MYH9, PARN, AT1A1, SIR2, SNRK, ZDHC5, CC50A, AMOT, AGAP3, MARK1, Q8VHM5, FLNC, SFPQ, CPIN1, WDR13, BACH, S12A5, RAB14, ACLY, MIC25, ATPG, DDX1, SH3L3, UBAP2, NCOA5, CSDE1, FRS3, ZFN2B, DLG2, PTBP2, SRGP1, TMLH, DYST, SYUB, ELOV6, ALS2, TADBP, TBB6, CLIP1, LRC59, K2C5, UBXN1, SIR1, SPRE1, PAWR, MED1, MEP50, STML2, UBP11, NONO, RRAGC, VMA5A, MAOM, DCTN2, NEUA, DDAH2, DNJA3, TRXR3, RB6I2, SRRT, DSRAD, Q99NC2, RIMS1, ANR17, NU155, NTRI, RRBP1, ZN318, TRI33, ATP5L, RL17, GLOD4, DUT, SDHB, GLRX3, IFM3, NECP1, OCAD1, RRP44, TBB2B, DDAH1, YIF1B, ROA0, NIP7, MPPB, CYBP, RL11, TECR, CHTOP, SERB1, QCR1, NNRD, GARS, TOM70, RS19, SYRC, CNDP2, TMEDA, ODO2, DLGP1, TBB4A, IDH3A, IPYR, RL37, FIP1, TIM50, EF1G, RM17, GSDMD, DDA1, F135B, TM263, CNN3, PLIN3, PGAM1, XRN2, MYPT1, DJC10, KC1D, GNAI3, PUR6, S38A3, NDUBA, CRIP2, TSC1, RAI14, NBEA, TCF20, SORC2, DPYL5, TBB3, RBP2, ARHG7, RTN3, SPN90, RBCC1, PSMG2, DDX24, CLD12, PALLD, ELF2, TMOD3, NUDT3, COPB, NUP50, DDX21, TULP4, FLII, RPF2, CCG3, TBA8, IQGA1, NECT1, ADRM1, FMN2, PALS1, DCLK1, BAG3, CUL3, MINK1, REEP6, TRXR1, SYGP1, SON, APBB1, DREB, SPY2, MACF1, ULK2, ZBP1, TOM40, ADDA, GOGA5, DNJB1, MAP1A, PCLO, GAB1, RIPK3, NPAS3, SH2D3, NUBP2, ZEB2, SYT7, DEST, TEBP, SRS10, RPGR, PR40A, KHDR3, TPSN, CDYL, KAD2, TEN1, PDC6I, CHIP, IF4H, COR1B, COR1C, TNIP1, GANP, ARC, MPP2, SHAN1, VAPA, GSK3B, DEMA, E41L3, JIP1, GBP2, CAD20, P5CS, LAT1, DYR1B, MD2L1, SAE2, APCL, SYVC, MTMR1, MECP2, E41L1, SUCB1, HDAC6, GRIA4, HOME1, OSB10
Species: Mus musculus, Rattus norvegicus
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Zaro BW, Yang YY, Hang HC, Pratt MR. Chemical reporters for fluorescent detection and identification of O-GlcNAc-modified proteins reveal glycosylation of the ubiquitin ligase NEDD4-1. Proceedings of the National Academy of Sciences of the United States of America 2011 108(20) 21540332
Abstract:
The dynamic modification of nuclear and cytoplasmic proteins by the monosaccharide N-acetyl-glucosamine (GlcNAc) continues to emerge as an important regulator of many biological processes. Herein we describe the development of an alkynyl-modified GlcNAc analog (GlcNAlk) as a new chemical reporter of O-GlcNAc modification in living cells. This strategy is based on metabolic incorporation of reactive functionality into the GlcNAc biosynthetic pathway. When combined with the Cu(I)-catalyzed [3 + 2] azide-alkyne cycloaddition, this chemical reporter allowed for the robust in-gel fluorescent visualization of O-GlcNAc and affinity enrichment and identification of O-GlcNAc-modified proteins. Using in-gel fluorescence detection, we characterized the metabolic fates of GlcNAlk and the previously reported azido analog, GlcNAz. We confirmed previous results that GlcNAz can be metabolically interconverted to GalNAz, whereas GlcNAlk does not, thereby yielding a more specific metabolic reporter of O-GlcNAc modification. We also used GlcNAlk, in combination with a biotin affinity tag, to identify 374 proteins, 279 of which were not previously reported, and we subsequently confirmed the enrichment of three previously uncharacterized proteins. Finally we confirmed the O-GlcNAc modification of the ubiquitin ligase NEDD4-1, the first reported glycosylation of this protein.
O-GlcNAc proteins:
E9QA16, EF2K, PRDX4, GSTO1, GSH0, DHX15, NOTC2, BCAT2, IMPA1, FHL2, S10A4, ANXA2, RIR1, HS90A, HME2, NUCL, ANXA1, EF1A1, CADH3, CATB, RLA0, H12, FGFR1, TPIS, COF1, VIME, CBX3, IMDH2, FKBP4, PRDX1, CAP1, ALD2, NEDD4, CAPZB, CRKL, LMNA, ROA1, ADT2, KPYM, HMGA2, RL10A, TB182, 1433E, RS3, RS27A, IF5A1, RS12, ACTS, TCPB, SGCG, CDS1, TBB5, XDH, CAPR2, CNN2, FOXO1, NB5R4, PRC2C, MILK2, FUBP2, PRRC1, SRC8, MYL6, VDAC2, VDAC1, RBBP7, ASNS, LAP2A, HCFC1, SERA, TIF1B, NUP62, ABCG1, G3PT, NOP58, TPM4, Q6NVA3, SYNE2, ACTN1, MBB1A, OTUB1, LPP, FLNA, BRI3B, LACC1, SYEP, EIF3B, MATR3, ABLM1, ERF3A, MYH9, CPIN1, WBP11, TRXR3, NPM3, CYBP, GARS, EF1G, CALR3, CP4V2, PUR6, ANM1, DDX21, DEST, CDYL, KHDR2, VAPA, TAGL2, SAE2, TRIP6, TULP1
Species: Mus musculus, Homo sapiens
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