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Luo Y, Wang Y, Tian Y, Zhou H, Wen L. "Two Birds One Stone" Strategy for the Site-Specific Analysis of Core Fucosylation and O-GlcNAcylation. Journal of the American Chemical Society 2023 37340703
Abstract:
Core fucosylation and O-GlcNAcylation are the two most famous protein glycosylation modifications that regulate diverse physiological and pathological processes in living organisms. Here, a "two birds one stone" strategy has been described for the site-specific analysis of core fucosylation and O-GlcNAcylation. Taking advantage of two mutant endoglycosidases (EndoF3-D165A and EndoCC-N180H), which efficiently and specifically recognize core fucose and O-GlcNAc, glycopeptides can be labeled using a biantennary N-glycan probe bearing azido and oxazoline groups. Then, a temperature-sensitive poly(N-isopropylacrylamide) polymer functionalized with dibenzocyclooctyne was introduced to facilitate the enrichment of the labeled glycopeptides from the complex mixture. The captured glycopeptides can be further released enzymatically by wild-type endoglycosidases (EndoF3 and EndoCC) in a traceless manner for mass spectrometry (MS) analysis. The described strategy allows simultaneous profiling of core-fucosylated glycoproteome and O-GlcNAcylated glycoproteome from one complex sample by MS technology and searching the database using different variable modifications.
O-GlcNAc proteins:
LCE6A, RBM47, HFM1, SMCO3, SBNO1, ODAD3, CNOT1, RCCD1, GLTD2, AGAP5, CX049, PDLI1, TAF4, ABLM1, DVL1, HGS, SC16A, NPC1, LAMA5, TET3, IF4G3, E41L2, AKAP8, PLIN3, MAFK, OPHN1, MITF, OBSL1, ANR17, ENTP6, NCOR1, ERLN1, JERKY, MYCB2, WDHD1, CBPD, TOX4, AGFG2, SC24B, PCNT, BAG3, DDAH2, CLPT1, AACT, LMNA, FINC, FETUA, GCR, KITH, HSPB1, RPN1, RLA2, ITB1, K1C18, ENOA, CATD, TBB5, TACD2, LYAG, BIP, LAMC1, HSP7C, DMD, MPRI, SKI, GILT, GLU2B, ENPL, RSMB, PO2F1, PVR, ZEP1, DPEP1, CBPE, ATF7, SON, ATF1, ITIH2, FST, ICAL, FGF7, CD9, CBL, ITA6, PTPRB, COF1, GATA3, PSA4, PEBP1, CLIP1, ZEP2, GLPK, ELF1, CD68, GPC1, HRH1, IRS1, NU214, SRP14, NUP62, ETFB, LICH, TXLNA, STAT3, MATR3, SSRA, GATA4, MMP13, 5HT3A, NOTC1, YAP1, RFX5, FAS, CDK8, CENPF, NU153, SEPP1, EMD, BCAM, HCFC1, SPHM, ARSD, AGFG1, NUP98, PTTG, RAD, AF17, DSRAD, ITA1, IF6, STAR6, ACTB, HNRPK, H4, RL40, CXAR, GPC5, FOXK1, PGBM, SPTB2, FOXK2, IF4G1, NOTC2, TLE3, PTN12, MTG8, ZO1, LRP1, RGS1, CD47, EP300, AHNK, TROAP, BPTF, NFIA, HYAL2, LMAN2, FOXC1, MB211, OS9, TUSC3, ROCK1, ASAH1, RIPK1, ASPP2, CDK13, SCRB2, VEZF1, DSG2, UBP2L, GIT2, PUM1, RRP1B, NCOA6, MEF2D, CHD4, NUMA1, R3HD1, RCN1, RBMS2, TAF1C, SF01, JHD2C, ELF2, TAB1, HERC1, ZFHX3, ZYX, ADRM1, CCDC6, SNPC1, MA2A1, YC018, QSER1, AAK1, P3H1, GNPTA, RABL6, TB10B, LUZP6, PRC2B, WIPI1, DCA10, HP1B3, ZN362, ZEP3, ZC3HD, UBR4, RHG21, UBAP2, RPRD2, DNAI4, TASO2, RN123, PCX4, ARID2, FTM, BICRL, SCAR3, GRHL2, NIPBL, LIN54, NFRKB, ZC3HE, LCN15, CREL2, IGS10, GGYF2, NBEL2, SRCAP, K0408, UBN2, BACHL, KDM3B, PARPT, RGPD4, POGZ, MAVS, EMSY, RAI1, I2BP2, ABCAC, ZFHX4, LUZP1, FRAS1, RB6I2, AHNK2, S22A9, TEX2, MGAP, SULF2, ANKH1, SUGP1, HYCC2, MILK2, CC116, PHAR4, K319L, ASPM, RPTOR, SYNPO, GALT4, MFSD9, SLAI1, CC168, TNR6A, PHC3, VP37A, SYNE1, PLBL2, TIP, CC110, TEX47, TBC15, STT3B, SPP2B, MAGC3, DYH5, PO210, GEMI5, PIGO, F222B, F151A, LMO7, P66B, MYO3B, GBF1, NICA, TM131, ZN592, LAR4B, GSLG1, GPKOW, LPP, TTC28, PF21A, RBM33, GWL, TONSL, PDLI5, VCIP1, ZFR, EP400, CH048, CI072, NOL4L, RBM14, GBP4, CDK15, PHF12, CIC, MED15, G3ST4, FNBP1, MINT, HTF4, EYA3, ARI3A, H2A1J, GDF15, DPH2, BCL7B, TM2D3, PELO, DIDO1, TRAIP, RBM4, CLC7A, UBE2O, PEG3, SP130, BRD8, I2BPL, EPC1, ADNP, RM46, NELFA, WNK1, ZHX3, SDS3, MLXIP, RC3H2, MUC5B, TANC2, CHD8, CELR2, APMAP, PDLI7, RBM12, STAU2, GPTC2, TAB2, CDK12, PTTG3, FLRT1, CRIM1, DAPLE, IBTK, RBM27, KANL3, RERE, SE1L1, LIMD1, TCF20, DPP2, BAP29, S30BP, LCAP, BTNL2, SIX4, POMT2, INT6, MRTFB, NOTC3, ATS5, BSN, SCAF8, ANR26, SHAN2, SRRM2, CTND2, SCML2, ZN652, ZN281, STRAP, VPP2, PRC2C, NCOR2, DC1L1, STON1, S23IP
Species: Homo sapiens
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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, 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, SERB1, ABCF1, Q8TDJ5, Q8WWH9, AGRV1, TCPW, TFG, STAM1, SNR40, VPS35, SIN3A, NIBA2, PSB7, TSNAX, DHRS6, 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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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,