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Lin CH, Liao CC, Wang SY, Peng CY, Yeh YC, Chen MY, Chou TY. Comparative O-GlcNAc Proteomic Analysis Reveals a Role of O-GlcNAcylated SAM68 in Lung Cancer Aggressiveness. Cancers 2022 14(1) 35008409
Abstract:
O-GlcNAcylation is a reversible and dynamic post-translational protein modification catalyzed by O-GlcNAc transferase (OGT). Despite the reported association of O-GlcNAcylation with cancer metastasis, the O-GlcNAc proteome profile for cancer aggressiveness remains largely uncharacterized. Here, we report our comparative O-GlcNAc proteome profiling of two differentially invasive lung adenocarcinoma cell lines, which identified 158 down-regulated and 106 up-regulated candidates in highly invasive cells. Among these differential proteins, a nuclear RNA-binding protein, SAM68 (SRC associated in mitosis of 68 kDa), was further investigated. Results showed that SAM68 is O-GlcNAcylated and may interact with OGT in the nucleus. Eleven O-GlcNAcylation sites were identified, and data from mutant analysis suggested that multiple serine residues in the N-terminal region are important for O-GlcNAcylation and the function of SAM68 in modulating cancer cell migration and invasion. Analysis of clinical specimens found that high SAM68 expression was associated with late cancer stages, and patients with high-OGT/high-SAM68 expression in their tumors had poorer overall survival compared to those with low-OGT/low-SAM68 expression. Our study revealed an invasiveness-associated O-GlcNAc proteome profile and connected O-GlcNAcylated SAM68 to lung cancer aggressiveness.
O-GlcNAc proteins:
A0A024R7P5, A0A024R9E2, A0A087WWU8, A0A0A0MTS7, SHOT1, A8K3C3, A8K9J7, B7Z2Z8, B7Z596, D3DS63, E7ETM0, E7EVA0, F8VR77, G3V1C3, H0YN18, H3BPE1, K7ERG4, PSD12, DFFA, PLOD2, PSDE, BIN1, TCRG1, ML12B, HGS, HNRDL, RPAC1, P4HA2, HNRPR, PLRG1, ZN207, BUB3, ACTN4, KDM1A, PLOD3, CPNE3, FLNB, NU155, GLRX3, MTA2, SC31A, UBE4B, TFR1, ANXA1, HSPB1, ITB1, DCUP, GELS, ENOA, NPM, TPM3, LDHB, ANXA2, TBB5, HNRPC, TPM2, ANXA6, 4F2, VIME, ANXA5, RSSA, ENOG, TPM1, PARP1, UBB, UBC, CH60, ACADM, G6PD, PCNA, KCRB, KCRU, ACTN1, XRCC6, EF2, KAP2, SYDC, AMPN, EZRI, NAGAB, HMGA1, ML12A, AOC1, ICAL, VATB2, FLNA, OSBP1, UBA1, GCSH, PSA1, PSA3, SYVC, TPP2, CLIP1, HNRH3, KINH, HSP74, RADI, MYH9, MYH10, ACTN2, ADDA, FUS, MYH11, RL4, ODO2, VATA, CAP2, GARS, MSH2, PRS6B, UBP5, RS9, MAP1B, IQGA1, KC1A, NASP, FAS, SYAC, NU153, HDGF, ACLY, SYYC, RD23A, PSMD4, TERA, EIF3B, IF6, PSA6, RS3A, HNRPK, 1433G, PP1B, PRS8, RL7A, PP2AB, RS6, RL10A, RS27A, RL40, 2ABA, TPM4, EF1A1, GTF2I, RAE1L, HNRPU, SPTB2, EWS, PLCB3, FKBP4, IF4G1, SSBP, 1433F, PUR1, PRDX1, KHDR1, ACTN3, PP2BA, ILF2, ACACA, CBX3, G3BP1, EIF3I, DC1I2, ROCK1, HDAC1, CUL1, NACA, SPTN1, SMC1A, GANAB, PSME4, SYK, PLEC, PP1R7, SC23A, SC23B, TSN, CIP4, MARE1, DDB1, CART, RBBP7, ACTBL, ST1C3, P4R3A, Q6IPH7, C2D1A, POTEE, SND1, CYFP1, MON2, MYH14, CAND1, ABCA7, LRC47, THMS1, CPSF7, GT251, PAIRB, ABCF1, Q8TDJ5, Q8WWH9, AGRV1, TCPW, TFG, STAM1, SNR40, VPS35, SIN3A, NIBA2, PSB7, TSNAX, BDH2, RBM4, XRN2, SPTN4, SLK, MYG1, XPP1, UGGG1, CPSF2, NAGK, NUDT5, PRP19, UBQL1, PACN2, SNX6, NCKP1, HYOU1, LSM4, SNX5, S23IP, V9HVZ7, V9HW77
Species: Homo sapiens
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Xu S, Zheng J, Xiao H, Wu R. Simultaneously Identifying and Distinguishing Glycoproteins with O-GlcNAc and O-GalNAc (the Tn Antigen) in Human Cancer Cells. Analytical chemistry 2022 35132862
Abstract:
Glycoproteins with diverse glycans are essential to human cells, and subtle differences in glycan structures may result in entirely different functions. One typical example is proteins modified with O-linked β-N-acetylglucosamine (O-GlcNAc) and O-linked α-N-acetylgalactosamine (O-GalNAc) (the Tn antigen), in which the two glycans have very similar structures and identical chemical compositions, making them extraordinarily challenging to be distinguished. Here, we developed an effective method benefiting from selective enrichment and the enzymatic specificity to simultaneously identify and distinguish glycoproteins with O-GlcNAc and O-GalNAc. Metabolic labeling was combined with bioorthogonal chemistry for enriching glycoproteins modified with O-GlcNAc and O-GalNAc. Then, the enzymatic reaction with galactose oxidase was utilized to specifically oxidize O-GalNAc, but not O-GlcNAc, generating the different tags between glycopeptides with O-GlcNAc and O-GalNAc that can be easily distinguishable by mass spectrometry (MS). Among O-GlcNAcylated proteins commonly identified in three types of human cells, those related to transcription and RNA binding are highly enriched. Cell-specific features are also revealed. Among glycoproteins exclusively in Jurkat cells, those involved in human T-lymphotropic virus type 1 (HTLV-1) infection are overrepresented, which is consistent with the cell line source and suggests that protein O-GlcNAcylation participated in the response to the virus infection. Furthermore, glycoproteins with the Tn antigen have different subcellular distributions in different cells, which may be attributed to the distinct mechanisms for the formation of protein O-GalNAcylation.
O-GlcNAc proteins:
RBM47, E2F8, SBNO1, CNOT1, HMX3, BTBDB, RHG32, P121C, PDLI1, SNP23, PSMD9, TAF4, ARI1A, ABLM1, STX16, HGS, MYPT1, SC16A, SR140, SET1A, FYB1, TIF1A, PPM1G, SHIP2, EIF3D, NUP42, KDM6A, TET3, SI1L1, DC1L2, HNRPR, PRPF3, TPD54, E41L2, ZN207, BUB3, AKAP8, ZNRD2, MYPT2, GANP, HNRPQ, DIAP1, PLIN3, MAFK, TBL1X, MITF, N4BP1, ZC11A, T22D2, PP6R2, ANR17, BCAS1, NCOR1, SPAG7, TIPRL, SPF30, TOX4, TOX, PCF11, AGFG2, ZFPL1, KIF4A, SC24A, SC24B, CNOT4, ASML, M4K4, BPNT1, PX11B, CHK2, LMNA, GLPA, TFR1, ALDOA, GCR, HSPB1, GNAI2, RLA1, RLA2, RLA0, K1C18, K2C8, RB, CATD, SYEP, PTPRC, VIME, GSTP1, HMGB1, ROA1, ATX1L, DERPC, ZN865, TPR, LAMP2, EF2, PLSL, PLST, GLU2B, HCLS1, PO2F1, RAC2, ATF2, ZEP1, TFE2, MUC1, CREB1, JUNB, ATF7, PTN2, DDX5, SON, ATF1, CSK22, NFKB1, FLNA, PUR2, RFX1, CBL, COF1, PTBP1, ARNT, DCK, PYR1, MAP4, CALX, 3MG, PRDX6, CDC27, AMRP, CLIP1, ZEP2, HNRH1, 1433S, ELF1, LSP1, PTN7, IRS1, ADDA, NU214, CUX1, TXLNA, MLH1, ECHA, IF2G, HNF4A, LAP2B, GPDM, RANG, KI67, CRKL, CAPZB, RFX5, SOX2, CAMLG, NASP, FAS, CDK8, SRP09, YLPM1, NU153, RBP2, TAF6, EMD, LRBA, PAPOA, HCFC1, HDGF, AGFG1, HNRPF, HXK2, NUP98, ATX1, RD23B, AF10, AF17, DSRAD, FOXA1, HNRH2, NU107, TPIS, PSME3, TPM4, F193A, GTF2I, PHC1, PRKDC, MAP1A, SARNP, FOXK1, FBLN2, FAM3A, EM55, NFKB2, HNRPU, SPTB2, FOXK2, RUNX1, FLI1, SATB1, SP2, MP2K1, NUCB1, KMT2A, IF4G1, TLE3, TLE4, KPCT, PSME1, GABPA, PRDX1, ACK1, AHNK, IFFO1, GALT2, SRBP2, TROAP, BPTF, TP53B, CBX3, NFAC2, PICAL, CUL4B, ASPP2, NFYC, CDK13, VEZF1, UBP2L, SRC8, CAPR1, LAGE3, PUM1, MDC1, EPN4, RRP1B, NCOA6, GSE1, UBP10, 2A5D, MEF2D, LASP1, NUMA1, CND1, TEBP, PCBP1, RBMS2, SF3A1, TSN, SF01, MED1, TRIP6, ELF2, TAB1, ZFHX3, ZYX, ADRM1, DPYL2, TAF9, MAPK3, CSPP1, PDS5A, QSER1, AAK1, LRRF1, VP26B, ACSF3, TPRN, CRTC2, PAN3, YIF1B, PRC2B, CEP78, ZN362, FKB15, LRIF1, CAF17, UBAP2, NT5D1, AHDC1, LYRM7, RPRD2, ZN318, TASO2, TBC9B, ARID2, C19L1, ABLM2, TWF2, GRHL2, CPZIP, NIPBL, LIN54, ZCHC8, C2D1A, SCYL2, NFRKB, RSBNL, MDEAS, ZC3HE, LARP1, SAMD1, FIP1, CRTC3, SAS6, MCAF1, BCOR, GGYF2, NBEL2, CO039, SRCAP, UBN2, TM1L2, ASXL2, SPT6H, MEPCE, BOP, KDM3B, ERMP1, TRM1L, ZCCHV, KANL1, POGZ, ZFY16, NUFP2, MAVS, EMSY, RAI1, I2BP2, SRGP1, RHG30, SH3R1, HUWE1, YTHD3, GALT7, LYRIC, BCL9L, CASZ1, TSYL5, DDX42, CACL1, P66A, I2BP1, VRK3, FOXP4, ARI3B, TEX2, MGAP, ANKH1, SUGP1, MILK2, ERF3B, K2013, PHAR4, XRN1, ZN687, FNBP4, ARFG1, ENAH, NHLC2, AVL9, XXLT1, GOLM1, TXND5, PAIRB, CHSTE, SLAI1, TNR6A, PHC3, SP20H, VP37A, KMT2C, ARI1B, KNL1, NEDD1, ALMS1, PREX1, DLG5, GEMI5, PIGO, UBS3B, WIPF2, FRS2, PDC6I, ZFN2B, TPC12, SEN15, PCNP, LMO7, ATX2L, CSKI2, PSPC1, P66B, GBF1, SMG7, RTF1, TOPB1, PHF3, MAML1, TTC9A, PRCC, RREB1, CBP, DDX17, SEM4D, ARHG1, GPKOW, FUBP2, LPP, TTC28, PF21A, FAF2, ESS2, EDC3, A7L3B, P121A, PDLI5, FUBP3, VCIP1, PDLI2, Z512B, ZFR, EP400, PRRC1, NOL4L, RBM14, PURB, NACC1, CIC, MED15, NUDC1, SIN3A, AEDO, MINT, HTF4, CNN2, RGPD5, ATX2, HCD2, S29A1, ARI3A, SH3G1, TRIR, DPH2, MGME1, ERP44, ESYT1, CCM2, CNPY3, WAC, DIDO1, HGH1, MMTA2, PAXX, NTM1A, RBM4, SGPP1, HEMGN, HDHD5, YTHD1, FTO, CEP44, BC11B, PITH1, SP130, BRD8, RGAP1, I2BPL, ADNP, DHX36, FOXP1, CENPH, WNK1, E41L1, ZHX3, YTDC2, RANB3, PHAX, ECT2, CNO10, MLXIP, PKHA5, PKHA1, RC3H2, LY9, RDH14, TAF9B, NCOA5, TANC2, TNR6C, CHD8, SDF2L, ARFG3, UBN1, RTN4, PDLI7, CHSTC, STRN4, PNO1, BMP2K, RBM12, STAU2, TXLNG, PNPO, CARF, TAB2, TMOD3, CDK12, F120A, HPBP1, ITSN2, CNOT2, CHMP5, VAPA, CAMP3, RBM27, KANL3, RERE, ZN219, SE1L1, STAP2, LIMD1, TCF20, SEPT9, UBQL2, TRPS1, S30BP, NRBP, EI2BD, SIX4, APC7, TASOR, GMEB2, PARP4, MA1B1, ACINU, ZHX1, CDV3, MRTFB, ZBT21, YETS2, HECD1, PKCB1, SCAF8, PP6R1, TRI33, TNR6B, ZC3H4, SHAN2, SRRM2, CTND2, SCML2, ZN148, T3JAM, VDAC3, AKAP2, DDX52, NOP58, GIT1, ZN281, SIT1, SALL2, ARIP4, CRBG1, HYOU1, KLF12, PRC2C, YTHD2, CD2AP, TNPO3, SRPRB, TSSC4, NUBP2, HCFC2, FHOD1, NCOR2, GMEB1, NCOA3, S23IP
Species: Homo sapiens
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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 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, PAIRB, 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 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, GLCM, 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,