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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, HCFC1, NEK4, AGFG1, NUP98, INHBC, CADH6, F193A, RT36, RT34, SARNP, LACTB, COG7, FOXK1, DAB2, PLIN5, SPTB2, SP2, NRG1, IF4G1, K1C17, TLE1, TLE3, 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, BZW2, CABIN, NCOA3, S23IP, U3KPZ7
Species: Homo sapiens
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Berthier A, Vinod M, Porez G, Steenackers A, Alexandre J, Yamakawa N, Gheeraert C, Ploton M, Maréchal X, Dubois-Chevalier J, Hovasse A, Schaeffer-Reiss C, Cianférani S, Rolando C, Bray F, Duez H, Eeckhoute J, Lefebvre T, Staels B, Lefebvre P. Combinatorial regulation of hepatic cytoplasmic signaling and nuclear transcriptional events by the OGT/REV-ERBα complex. Proceedings of the National Academy of Sciences of the United States of America 2018 115(47) 30397120
Abstract:
The nuclear receptor REV-ERBα integrates the circadian clock with hepatic glucose and lipid metabolism by nucleating transcriptional comodulators at genomic regulatory regions. An interactomic approach identified O-GlcNAc transferase (OGT) as a REV-ERBα-interacting protein. By shielding cytoplasmic OGT from proteasomal degradation and favoring OGT activity in the nucleus, REV-ERBα cyclically increased O-GlcNAcylation of multiple cytoplasmic and nuclear proteins as a function of its rhythmically regulated expression, while REV-ERBα ligands mostly affected cytoplasmic OGT activity. We illustrate this finding by showing that REV-ERBα controls OGT-dependent activities of the cytoplasmic protein kinase AKT, an essential relay in insulin signaling, and of ten-of-eleven translocation (TET) enzymes in the nucleus. AKT phosphorylation was inversely correlated to REV-ERBα expression. REV-ERBα enhanced TET activity and DNA hydroxymethylated cytosine (5hmC) levels in the vicinity of REV-ERBα genomic binding sites. As an example, we show that the REV-ERBα/OGT complex modulates SREBP-1c gene expression throughout the fasting/feeding periods by first repressing AKT phosphorylation and by epigenomically priming the Srebf1 promoter for a further rapid response to insulin. Conclusion: REV-ERBα regulates cytoplasmic and nuclear OGT-controlled processes that integrate at the hepatic SREBF1 locus to control basal and insulin-induced expression of the temporally and nutritionally regulated lipogenic SREBP-1c transcript.
O-GlcNAc proteins:
A4D111, POTEF, A5GZ75, AXA2L, P121C, A9Z0R7, EIFCL, C3UMV2, F1JVV5, I6TRR8, MYO1C, IF2B3, DDX3X, TCRG1, OPLA, XPO1, SC16A, SET1A, OGT1, EIF3D, DDX3Y, DHX15, PRP4, SERA, PSMD3, HNRPR, ACTN4, MYO1B, AKAP8, HNRPQ, UGDH, USO1, WDR1, ANR17, GGCT, LX12B, FLNB, PR40A, SF3B1, SPB7, NU155, KRT38, SC24D, GLSK, SC31A, ELP1, SMC2, AGM1, UTS2, BAG4, SC24A, SC24B, AP2A1, LDHA, AL1A1, PGK1, A2MG, CO3, CYTA, KV117, IGHG1, IGHA1, APOE, APOC2, FIBG, TFR1, TRFE, CATA, ALDOA, TBB4A, G3P, HSPB1, RPN1, RPN2, AT1A1, ARGI1, ALDH2, S10A8, ADT2, GELS, ATPB, APOA4, ENOA, PYGL, G6PI, TPM3, PDIA1, CATD, ANXA2, CAN1, TBB5, HS90A, SP1, CO1A2, HS90B, PO2F2, GSTP1, VILI, ANXA4, PARP1, LKHA4, ATX1L, POTEI, UBB, UBC, SAA2, HS71A, HS71B, IGG1, TBA3C, TBA3D, THIO, CH60, BIP, HSP7C, PYGB, PYGM, G6PD, PYC, C1TC, NFH, IMDH2, XRCC6, XRCC5, AT1A3, EF2, PDIA4, P4HA1, ENOB, GFAP, ENPL, IDE, PO2F1, HNRPL, PLAK, DESP, AT2A2, HSP76, DDX5, LEG3, TCPA, RL7, VINC, E2AK2, ITIH2, ANXA7, HNF1A, FILA, CD11B, FLNA, VDAC1, TGM2, PUR2, UBA1, NDKB, TGM1, EST1, SFPQ, SAHH, MCM3, ATPA, PTBP1, SYVC, ABCD3, GRN, TKT, SPB3, AL4A1, PDIA3, KPYR, RPB2, AKT1, PUR9, HNRH1, CASPE, 1433S, S10AB, PRDX2, MCM4, MCM7, HS71L, CTNB1, IRS1, GDE, MYH9, FUS, SPB5, NUP62, TALDO, GRP75, CAPG, TCPZ, STAT3, MDHC, MDHM, ECHA, GARS, SYIC, HUTH, LPPRC, MATR3, MSH2, VDAC2, SYQ, LEG7, COPD, SPB4, TCPE, AL9A1, LMAN1, FMO5, TCPG, SYAC, RBM25, KLK7, DYN2, TCPQ, TCPD, RAB7A, HCFC1, KS6A3, HNRPM, HXK2, CAZA1, NUP98, ACLY, COPB, COPA, SC24C, SYRC, SYYC, UBP14, HSP72, P5CS, XPO2, TERA, MTP, AF17, PSA, HNRH2, EIF3B, SYMC, NU107, EPIPL, TPIS, ACTB, IF4A1, HNRPK, 1433G, PRS4, ACTA, H4, RS27A, RL40, 1433Z, RACK1, ACTG, ACTH, ACTC, ACTS, TBA1B, TBA4A, TBB4B, PRKDC, DCD, VIGLN, CLH1, HNRPU, FABP5, MSHR, EWS, SEMG2, DSG1, SP3, PLOD1, EF1A2, GFPT1, PRDX1, KHDR1, TGM3, DHX9, LG3BP, DSC1, ILF3, TRAP1, PAK2, PSMD2, PABP4, PICAL, PKP1, BLMH, SNTB1, TBB2A, VEZF1, TRI29, UBP2L, LY6D, SRC8, PDIA5, HS902, EPN4, SMC1A, GANAB, MVP, PLEC, NONO, SC23A, SC23B, CDSN, JHD2C, CYTM, DPYL2, PCKGM, TKFC, Q53G76, Q58FF2, Q59EA0, ZN326, FILA2, UBAP2, XP32, RBM26, EF1A3, ARID2, TBA3E, POTEE, SBSN, FBX50, Q70T18, Q71E78, TBA1A, SND1, NUP54, MYH14, PEG10, PRP39, TAXB1, CAND1, CARM1, PRSR1, SPA12, ANKH1, ASXL1, NUP93, RDHE2, Q8N6B4, PDPR, TNR6A, COP1, PDC6I, POF1B, ATX2L, DDX1, BAP1, TFG, RBP56, EVPL, DDX17, RENT1, FUBP2, UBP7, NCLN, H2B1A, WNK4, ZC3HA, SCYL1, SPB12, GSDMA, VPS35, PHF12, CIC, STRBP, VAT1, NUP88, ATX2, CPNE1, TCPH, TBA1C, DIDO1, HNRL1, TBB2B, NUP58, ACTBM, TB182, SP130, WNK1, AGO3, MCCB, MOV10, TNR6C, S10AE, DD19A, ATD3A, TBA8, UGGG1, IF2B1, CALL5, RRBP1, NXF1, CMC2, PO2F3, AGO2, AGO1, Q9UL79, ACSL5, DD19B, TNR6B, CD11A, EIF3L, SYFA, KLK5, RTCB, WNK2, PKP3, HYOU1, SNX9, COPG1, IF2B2, S23IP
Species: Homo sapiens
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