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He J, Fan Z, Tian Y, Yang W, Zhou Y, Zhu Q, Zhang W, Qin W, Yi W. Spatiotemporal Activation of Protein O-GlcNAcylation in Living Cells. Journal of the American Chemical Society 2022 144(10) 35138101
Abstract:
O-linked N-acetylglucosamine (O-GlcNAc) is a prevalent protein modification that plays fundamental roles in both cell physiology and pathology. O-GlcNAc is catalyzed solely by O-GlcNAc transferase (OGT). The study of protein O-GlcNAc function is limited by the lack of tools to control OGT activity with spatiotemporal resolution in cells. Here, we report light control of OGT activity in cells by replacing a catalytically essential lysine residue with a genetically encoded photocaged lysine. This enables the expression of a transiently inactivated form of OGT, which can be rapidly reactivated by photo-decaging. We demonstrate the activation of OGT activity by monitoring the time-dependent increase of cellular O-GlcNAc and profile glycoproteins using mass-spectrometry-based quantitative proteomics. We further apply this activation strategy to control the morphological contraction of fibroblasts. Furthermore, we achieved spatial activation of OGT activity predominantly in the cytosol. Thus, our approach provides a valuable chemical tool to control cellular O-GlcNAc with much needed spatiotemporal precision, which aids in a better understanding of O-GlcNAc function.
O-GlcNAc proteins:
SBNO1, CNOT1, BACH, PSD11, PSD12, TAF4, CLIC1, EIF3F, IPO5, IF2B3, ARI1A, KMT2D, ANM5, PSA7, HAT1, HGS, MYPT1, XPO1, SC16A, SR140, SET1A, PUR4, NPC1, OGT1, HMGB3, PPM1G, EIF3D, EIF3H, P4HA2, SERA, PSMD3, PAPS1, MSI1H, IF4G3, E41L2, FOXO3, ZN207, BUB3, ACTN4, SYNC, SAHH2, KPRB, GANP, PEPL, OGA, PLOD3, IMA7, IF2P, DNJA2, MITF, CPNE3, CLU, PP6R2, CREST, ANR17, NCOR1, VP26A, CLN5, CSDE1, IDHC, SRP72, MTA2, TOX4, SC24D, PCF11, NFAT5, SC31A, AGFG2, SCAF4, SMC2, IPO7, PSMG1, SC24A, SC24B, EYA4, HS74L, TOM40, LDHA, PNPH, HPRT, PGK1, CAH2, ALDOA, ANXA1, G3P, IF2A, RLA1, RLA2, RLA0, JUN, LA, AGAL, KCRM, ENOA, PYGL, G6PI, LDHB, H10, ANXA2, TBB5, PROF1, APT, SYEP, HS90A, LAMB1, SP1, ANXA6, DAF, PFKAM, HS90B, ASNS, RS17, ANXA5, RSSA, GSTP1, HMGB1, PARP1, LKHA4, ALDOC, ATX1L, HS71B, RO60, PTPRF, THIO, HSP7C, EPB41, UMPS, G6PD, C1TC, ADHX, SRF, PRPS2, PABP1, PCNA, IMDH2, KCRB, PEPD, XRCC6, XRCC5, RINI, EF2, P4HA1, PLST, ACPH, GYS1, KPYM, PO2F1, SYDC, PLAK, ERF3A, NDKA, RS2, CBR1, CREB1, HSP76, PYRG1, DDX5, PFKAL, TCPA, RL35A, ARF4, RL7, RL17, PGAM1, DNLI1, NUCL, SPEE, CSK22, PSB1, FLNA, PIMT, PUR2, PUR6, UBA1, NDKB, RFX1, CBL, RS3, NFYA, SAHH, COF1, EF1B, MCM3, RS12, BRD2, PSA1, PSA2, PSA3, PSA4, MOES, DDX6, DNMT1, PAX6, U2AF2, RL13, SYTC, SYVC, EF1G, 1433T, ARNT, RL10, RFA1, APEX1, PYR1, MAP4, PSB6, PSB5, AMPL, TKT, RBMS1, EF1D, PRDX6, RL12, PEBP1, 2AAA, CDC27, NMT1, PURA2, PUR8, METK2, DNJA1, PUR9, 1433B, STIP1, PRDX2, ELF1, CGL, RL9, KINH, MCM4, MCM5, MCM7, HSP74, RL22, CBS, MYH9, MYH10, COPB2, FUS, DEK, PRS7, RL4, SRP14, TALDO, RS19, RL3, TCPZ, RL13A, MDHC, IF2G, CSK, GARS, SYIC, RS27, RANG, BAG6, NSF, RL27A, RL5, RL21, RL28, RS9, RS10, SYQ, RL29, ATPO, PPCE, COPD, TCPE, PIPNB, AL9A1, NASP, FAS, TCPG, SYAC, SYSC, PSB3, MCM2, YLPM1, RBM25, HINT1, GSK3A, GUAA, DNLI3, GDIB, SERPH, F10A1, RL14, TCPQ, TCPD, ANX11, PAPOA, SMCA4, HCFC1, SSDH, 6PGD, IMA1, AGFG1, HNRPF, THOP1, PPP5, ACLY, COPB, COPA, SC24C, SYRC, ATN1, SYYC, RD23B, ANAG, XPO2, TERA, NP1L1, PSA, EIF3B, ATPK, SYMC, TPIS, EIF3E, IF4A1, RS20, PRPS1, PSA6, CDC42, UBC12, UBE2N, ARP3, ARP2, ACTZ, CSN2, ABCE1, RS3A, RL26, RL15, RL27, 1433G, RS7, PRS8, RS8, RS15A, RS16, 1433E, RS23, RS18, RS13, RS11, RUXE, PRS10, RL7A, ERF1, RS4X, RL23A, RS6, RAN, RL23, UB2D2, RS24, RS25, RS26, RL30, RL10A, RL32, RL11, RL8, PPIA, RS27A, RAC1, AP2B1, 1433Z, RSMN, SUMO1, RL38, IF5A1, RACK1, YBOX1, EF1A1, TBA1B, CSK21, F193A, IF4G2, PHC1, TCPB, GSTO1, RL24, RL36A, ARF1, RL19, FOXK1, RBM10, CYC, CLH1, SPTB2, SET, FOXK2, CAP1, OTUD4, EWS, SP3, RL18A, FKBP4, RL6, KMT2A, IF4G1, TLE3, TLE4, 1433F, SRS11, EF1A2, GFPT1, EXOS9, SUH, GABPA, PRDX1, RL18, SRSF1, SSRP1, RBBP4, EP300, AP1B1, SFSWA, FOXC1, ACACA, CSN1, AIMP2, PSMD2, G3BP1, PABP4, EIF3I, SF3B2, PICAL, ULA1, CUL4B, FHL1, NACA, SPTN1, NFYC, CKAP5, EIF3A, UBP2L, TTL12, DYHC1, RCN2, CAPR1, RBM39, PUM1, EPN4, NCOA6, GSE1, MEF2D, ZN638, IMB1, NOLC1, NUMA1, PSMD6, SEPT2, R3HD1, BRD3, PA1B3, IPYR, TEBP, RCN1, PCBP1, PCBP2, SC23A, SF3A1, NCOA2, SF01, MED1, JHD2C, ELF2, TAB1, TBCE, VAS1, ZYX, SEPT7, ADRM1, CCDC6, PKN2, DDB1, CDC37, NRF1, FSCN1, RFX7, QSER1, QRIC1, TBB8, LARP7, TB10B, AMOT, TGO1, PRC2B, UBAP2, QSPP, RBM26, RPRD2, TASO2, TSH3, ARID2, LIN54, EDC4, SCYL2, NFRKB, ZC3HE, FIP1, MCAF1, BCOR, UBN2, LARP4, SPT6H, SND1, DDX46, CYFP1, KDM3B, ZCCHV, NUFP2, PLGT3, RAI1, RBBP6, SH3R1, HUWE1, YTHD3, CENPV, KAISO, KTN1, CAND1, RTTN, CARM1, PRSR1, P66A, SPA12, Z3H7A, ANKH1, SUGP1, CCAR1, PHC2, SMAP1, PHAR4, DCP1B, FNBP4, CPSF7, ARFG1, ENAH, SUMF2, PGLT1, SERB1, LS14A, TNR6A, ABCF1, NEDD1, WDR36, SMRC2, PO210, PDC6I, ATX2L, P66B, DDX1, SMG7, MAML1, HS105, LAR4B, GCN1, AN32B, TFG, CBP, RENT1, SMRC1, FUBP2, TNPO1, USP9X, NCLN, FERM2, FKB10, P5CR2, ISOC1, NMD3, EDC3, OTUB1, PDLI5, FUBP3, ZC3HA, EP400, PRRC1, RBM14, VPS35, CIC, MED15, SEC62, PSMD1, PARK7, EYA3, VAT1, SCAFB, EIF3C, ATX2, TS101, TCPH, ANM1, RNZ2, TBA1C, CNPY3, WAC, DIDO1, AN32E, TBB6, HNRL1, TBB2B, GNL3, THIC, RBM4, NAA15, YTHD1, WNK3, UNK, UBA5, BRD8, LMA2L, FOXP1, NELFA, PTN23, WNK1, AMPB, RPF2, GORS2, LRC40, MLXIP, MYG1, RISC, CYBP, RC3H2, TAF9B, NCOA5, CHD8, CELR2, DCP1A, PDLI7, SAR1A, SHLB2, MBNL1, SALL1, SYFB, PDS5B, OLA1, RBM12, DD19A, FANCI, LYAR, CARF, TAB2, UGGG1, CDK12, IF2B1, ITSN2, BICRA, CNOT2, RCC2, SYLC, RBM27, KANL3, ATX10, SAE1, SAE2, SUN2, SRP68, CHRD1, UBQL2, S30BP, PUF60, DACH1, SIX4, HOOK1, MRT4, NUP50, MRTFB, ZMIZ1, YETS2, HECD1, MYO6, PRP19, UBQL1, G3BP2, MAGD2, CSN3, SCAF8, TRI33, SRRM2, PA2G4, RUVB2, EIF3L, DRG1, OFUT2, E41L3, R3HD2, RRP44, NOP58, ZN281, LC7L2, SBDS, STRAP, RTCB, SALL2, TLN1, ARIP4, HYOU1, KLF12, ARI1, PRC2C, YTHD2, SP16H, SERC, GMEB1, ZHX2, S23IP
Species: Homo sapiens
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Liu J, Hao Y, Wang C, Jin Y, Yang Y, Gu J, Chen X. An Optimized Isotopic Photocleavable Tagging Strategy for Site-Specific and Quantitative Profiling of Protein O-GlcNAcylation in Colorectal Cancer Metastasis. ACS chemical biology 2022 17(3) 35254053
Abstract:
O-linked-β-N-acetylglucosamine (O-GlcNAc) glycosylation is a ubiquitous protein post-translational modification of the emerging importance in metazoans. Of the thousands of O-GlcNAcylated proteins identified, many carry multiple modification sites with varied stoichiometry. To better match the scale of O-GlcNAc sites and their dynamic nature, we herein report an optimized strategy, termed isotopic photocleavable tagging for O-GlcNAc profiling (isoPTOP), which enables quantitative and site-specific profiling of O-GlcNAcylation with excellent specificity and sensitivity. In HeLa cells, ∼1500 O-GlcNAcylation sites were identified with the optimized procedures, which led to quantification of ∼1000 O-GlcNAcylation sites with isoPTOP. Furthermore, we apply isoPTOP to probe the O-GlcNAcylation dynamics in a pair of colorectal cancer (CRC) cell lines, SW480 and SW620 cells, which represent primary carcinoma and metastatic cells, representatively. The stoichiometric differences of 625 O-GlcNAcylation sites are quantified. Of these quantified sites, many occur on important regulators involved in tumor progression and metastasis. Our results provide a valuable database for understanding the functional role of O-GlcNAc in CRC. IsoPTOP should be applicable for investigating O-GlcNAcylation dynamics in various pathophysiological processes.
O-GlcNAc proteins:
A0A0B4J203, A0A0C4DFX4, RBM47, E2F8, WDR27, SBNO1, CNOT1, P121B, P121C, H0YAE9, H0YHG0, H7C469, K7ELQ4, M0QZ24, PDLI1, HAX1, TAF4, BCL9, CAC1A, DDX3X, NFIB, PPP6, MA2B1, ARI1A, SOCS7, ABLM1, KMT2D, GBRD, RGRF2, TX1B3, HGS, MYPT1, SYN3, ZN609, TRI66, PDZD2, MAST4, SC16A, SET1A, CASC3, FOXP2, MOT4, P4HA2, ARPC5, CLOCK, MAFG, PER1, KDM6A, TET3, SI1L1, TGFI1, M3K7, MCA3, PRPF3, TPD54, SYNJ1, IF4G3, E41L2, WIPF1, FOXO3, TGM5, RNF13, SPY2, PLRG1, ZN207, AKAP8, CALU, ORC5, MYPT2, GANP, OGA, CCNT1, BUB1B, PLOD3, PLIN3, MOT2, MAFK, PQBP1, BRD4, TBL1X, PP1RB, NBN, MITF, SRGP2, N4BP1, ROCK2, PP6R2, CNOT3, ANR17, FLNB, NCOR1, SF3B1, REM1, CREG1, CRTAP, SYUG, CYTF, TOX4, TOX, SUN1, PCF11, AGFG2, UBE4B, CAC1H, SVIL, SC24A, SC24B, CNOT4, EYA4, ZMYM6, BAG3, LATS1, DDAH2, TXD12, ONEC2, CLPT1, ABL1, CRYAB, LMNA, TFR1, CATA, GBA1, FUCO, ALDOA, GCR, G3P, CPNS1, HSPB1, RLA2, RLA0, ITB1, K1C18, NPM, CATL1, CATB, MCR, BGLR, ITA5, NFIC, VIME, SNRPA, FGR, ATX1L, DERPC, ZN865, GLI2, MYBB, CLUS, PPAL, MPRI, PABP1, TPR, BMP3, SKIL, ENPL, PO2F1, PLAK, ATF2, ZEP1, RS2, TFE2, F261, ITB4, ZNF23, ZNF25, JUNB, ATF7, TPH1, DDX5, EGR1, SON, NELFE, ATF1, ATF6A, CADH2, ICAL, CSRP1, FLNA, RFX1, CBL, SFPQ, COF1, IF4B, GATA2, APC, DDX6, ARNT, MAP4, LYOX, HXD9, MZF1, CLIP1, 5HT1F, HXA11, ZEP2, ELF1, CTNB1, FBN1, ADDA, BASI, NU214, VGFR2, SRP14, NUP62, SYUA, VATA, CUX1, TXLNA, STAT3, LAP2A, EPS15, HELZ, MATR3, SSRA, SSRB, KI67, ATRX, MAP1B, YAP1, UTRN, STT3A, SC6A8, RFX5, SOX2, PRC2A, HSP13, NR2C2, NASP, CDK8, DHE4, YLPM1, NU153, RBP2, TAF6, MRE11, EMD, MXI1, MAP2, TOB1, PPT1, TCPQ, PAPOA, HCFC1, GDS1, AGFG1, CRIP2, NUP98, SMTN, SC24C, HIRA, ATX1, ATN1, AFAD, AF10, AF17, DSRAD, SEC13, NU107, ZN445, CSN2, RL37, WDR5, TIM10, F193A, RBM6, PITX1, IF4G2, PHC1, ADA17, KGD4, RL19, FOXK1, DAB2, RHG04, RBM10, HNRPU, SPTB2, FOXK2, RUNX1, MEF2A, SP2, SP3, PLOD1, KMT2A, TF65, IF4G1, NOTC2, TLE3, TLE4, PTN12, CALD1, MEF2C, P5F1B, GABPA, ZO1, ACK1, EP300, AHNK, FCHO2, HMGX3, SRBP2, FOXO1, ASPH, TROAP, BPTF, FSTL1, NFIA, DPYD, TP53B, FOXC1, ECH1, ROA0, DDX10, TBX2, GPS2, G3BP1, PABP4, ADAM9, PICAL, NAB1, SERC3, RIPK1, IQGA2, STIM1, CUL4B, ASPP2, CAC1S, RUNX2, NFYC, CDK13, TOB2, VEZF1, UBP2L, GIT2, SRC8, CAPR1, LAGE3, PUM1, MDC1, EPN4, TTLL4, RRP1B, NCOA6, GSE1, MEF2D, LASP1, MYPC3, ZN638, NUMA1, SART3, CND1, R3HD1, KIF14, WDR43, PLCL1, PLEC, NOMO1, NONO, RCN1, RYR3, KS6A1, RBMS2, TAF1C, SF01, MED1, JHD2C, TRIP6, T22D1, ELF2, TAB1, HERC1, NCOA1, VAS1, ZFHX3, ZYX, ADRM1, SYPL1, TAF9, DREB, DGKD, CGT, GEN, LY6K, RFX7, QSER1, AAK1, PRSR3, QRIC1, MA7D1, WDR72, TBRG1, TB10B, TPRN, FIL1L, SVEP1, AMOT, EPC2, CRTC2, PAN3, HS904, YIF1B, AG10A, IGS11, ZN628, BCORL, FIGN, K2026, SH319, TGO1, PRC2B, TOIP1, CEP78, P4R3B, HP1B3, CE170, ZN362, FKB15, AKND1, ZEP3, LRIF1, SWT1, RHG21, UBAP2, RBM26, DEP1A, OGRL1, AHDC1, F222A, RPRD2, RN220, ZN318, TASO2, ZMYM4, PAPD7, TENS2, KANK2, ARID2,