Wikipedia

Protein O-GlcNAc transferase

(Redirected from OGT (gene))

Protein O-GlcNAc transferase also known as OGT or O-linked N-acetylglucosaminyltransferase is an enzyme (EC 2.4.1.255) that in humans is encoded by the OGT gene.[5][6] OGT catalyzes the addition of the O-GlcNAc post-translational modification to proteins.[7][8][9][10][5][11]

OGT
Available structures
PDBOrtholog search: PDBe RCSB
Identifiers
AliasesOGT, HRNT1, O-GLCNAC, HINCUT-1, O-linked N-acetylglucosamine (GlcNAc) transferase, OGT1, MRX106, XLID106
External IDsOMIM: 300255; MGI: 1339639; HomoloGene: 9675; GeneCards: OGT; OMA:OGT - orthologs
Orthologs
SpeciesHumanMouse
Entrez

8473

108155

Ensembl

ENSG00000147162

ENSMUSG00000034160

UniProt

O15294

Q8CGY8

RefSeq (mRNA)

NM_003605
NM_181672
NM_181673
NM_025192

NM_001290535
NM_139144

RefSeq (protein)

NP_858058
NP_858059

NP_001277464
NP_631883

Location (UCSC)Chr X: 71.53 – 71.58 MbChr X: 100.68 – 100.73 Mb
PubMed search[3][4]
Wikidata
View/Edit HumanView/Edit Mouse

Nomenclature

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Other names include:

  • O-GlcNAc transferase
  • OGTase
  • O-linked N-acetylglucosaminyltransferase
  • Uridine diphospho-N-acetylglucosamine:polypeptide β-N-acetylglucosaminyltransferase

Systematic name: UDP-N-α-acetyl-d-glucosamine:[protein]-3-O-N-acetyl-β-d-glucosaminyl transferase

Function

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O-GlcNAc transferase
Identifiers
EC no.2.4.1.255
Databases
IntEnzIntEnz view
BRENDABRENDA entry
ExPASyNiceZyme view
KEGGKEGG entry
MetaCycmetabolic pathway
PRIAMprofile
PDB structuresRCSB PDB PDBe PDBsum
Search
PMCarticles
PubMedarticles
NCBIproteins

Glycosyltransferase

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OGT catalyzes the addition of a single N-acetylglucosamine through an O-glycosidic linkage to serine or threonine and an S-glycosidic linkage to cysteine[12][13] residues of nucleocytoplasmic proteins. Since both phosphorylation and O-GlcNAcylation compete for similar serine or threonine residues, the two processes may compete for sites, or they may alter the substrate specificity of nearby sites by steric or electrostatic effects. Two transcript variants encoding cytoplasmic and mitochondrial isoforms have been found for this gene.[6] OGT glycosylates many proteins including: Histone H2B,[14] AKT1,[15] PFKL,[16] KMT2E/MLL5,[16] MAPT/TAU,[17] Host cell factor C1,[18] and SIN3A.[19]

O-GlcNAc transferase is a part of a host of biological functions within the human body. OGT is involved in the resistance of insulin in muscle cells and adipocytes by inhibiting the Threonine 308 phosphorylation of AKT1, increasing the rate of IRS1 phosphorylation (at serine 307 and serine 632/635), reducing insulin signaling, and glycosylating components of insulin signals.[20] Additionally, O-GlcNAc transferase catalyzes intracellular glycosylation of serine and threonine residues with the addition of N-acetylglucosamine. Studies show that OGT alleles are vital for embryogenesis, and that OGT is necessary for intracellular glycosylation and embryonic stem cell vitality.[21] O-GlcNAc transferase also catalyzes the posttranslational modification that modifies transcription factors and RNA polymerase II, however the specific function of this modification is mostly unknown.[22]

Protease

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OGT cleaves Host Cell Factor C1, at one or more of 6 repeating 26 amino acid sequences. The TPR domain of OGT binds to the carboxyl terminal portion of an HCF1 proteolytic repeat so that the cleavage region is in the glycosyltransferase active site above uridine-diphosphate-GlcNAc [11] The large proportion of OGT complexed with HCF1 is necessary for HCF1 cleavage, and HCFC1 is required for OGT stabilization in the nucleus. HCF1 regulates OGT stability using a post-transcriptional mechanism, however the mechanism of the interaction with HCFC1 is still unknown.[23]

Structure

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The human OGT gene has 1046 amino acid residues, and is a heterotrimer consisting of two 110 kDa subunits and one 78 kDa subunit.[10] The 110 kDa subunit contains 13 tetratricopeptide repeats (TPRs); the 13th repeat is truncated. These subunits are dimerized by TPR repeats 6 and 7. OGT is highly expressed in the pancreas and also expressed in the heart, brain, skeletal muscle, and the placenta. There have been trace amounts found in the lung and the liver.[5] The binding sites have been determined for the 110 kDa subunit. It has 3 binding sites at amino acid residues 849, 852, and 935. The probable active site is at residue 508.[16]

The crystal structure of O-GlcNAc transferase has not been well studied, but the structure of a binary complex with UDP and a ternary complex with UDP and a peptide substrate has been researched.[11] The OGT-UDP complex contains three domains in its catalytic region: the amino (N)-terminal domain, the carboxy (C)-terminal domain, and the intervening domain (Int-D). The catalytic region is linked to TPR repeats by a translational helix (H3), which loops from the C-cat domain to the N-cat domain along the upper surface of the catalytic region.[11] The OGT-UDP-peptide complex has a larger space between the TPR domain and the catalytic region than the OGT-UDP complex. The CKII peptide, which contains three serine residues and a threonine residue, binds in this space. In 2021 a 5Å CryoEM analysis revealed the relationship between the catalytic domains[24] and the intact TPR regions confirming the dimer arrangement first seen in the TPR alone X ray structure. This structure supports an ordered sequential bi-bi mechanism that matches the fact that “at saturating peptide concentrations, a competitive inhibition pattern was obtained for UDP with respect to UDP-GlcNAc.”[11]

Mechanism of catalysis

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The molecular mechanism of O-linked N-acetylglucosamine transferase has not been extensively studied either, since there is not a confirmed crystal structure of the enzyme. A proposed mechanism by Lazarus et al. is supported by product inhibition patterns of UDP at saturating peptide conditions. This mechanism proceeds with starting materials Uridine diphosphate N-acetylglucosamine, and a peptide chain with a reactive serine or threonine hydroxyl group. The proposed reaction is an ordered sequential bi-bi mechanism.[11]

 
Figure 2: The proposed catalytic mechanism of O-GlcNAc transferase. It is an ordered sequential bi-bi mechanism with only one step, not including a proton transfer. The peptide is represented by a serine residue with a reactive hydroxyl group.[11]

The chemical reaction can be written as:

  1. UDP-N-acetyl-D-glucosamine + [protein]-L-serine → UDP + [protein]-3-O-(N-acetyl-D-glucosaminyl)-L-serine
  2. UDP-N-acetyl-D-glucosamine + [protein]-L-threonine → UDP + [protein]-3-O-(N-acetyl-D-glucosaminyl)-L-threonine

First, the hydroxyl group of serine is deprotonated by histidine 498, a catalytic base in this proposed reaction. Lysine 842 is also present to stabilize the UDP moiety. The oxygen ion then attacks the sugar-phosphate bond between the glucosamine and UDP. This results in the splitting of UDP-N-acetylglucosamine into N-acetylglucosamine – peptide and UDP. Proton transfers take place at the phosphate and histidine 498. This mechanism is spurred by OGT gene containing O-linked N-acetylglucosamine transferase. Aside from proton transfers the reaction proceeds in one step, as shown in Figure 2.[11] Figure 2 uses a lone serine residue as a representative of the peptide with a reactive hydroxyl group. Threonine could have also been used in the mechanism.

Inhibitors

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Many inhibitors of OGT enzymatic activity have been reported.[25] OGT inhibition results in global downregulation of O-GlcNAc. Cells appear to upregulate OGT and downregulate OGA in response to OGT inhibition.[26]

5S-GlcNAc

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Ac45S-GlcNAc is converted intracellularly into UDP-5S-GlcNAc, a substrate analogue inhibitor of OGT. UDP-5S-GlcNAc is not efficiently utilized as a donor sugar by OGT, possibly due to distortion of the pyranose ring by replacement of oxygen with sulfur.[26] As other glycosyltransferases utilize UDP-GlcNAc as a donor sugar, UDP-5S-GlcNAc has some non-specific effects on cell-surface glycosylation.[27]

OSMI

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OSMI-1 was first identified from high-throughput screening using fluorescence polarization.[27] Further optimization led to the development of OSMI-2, OSMI-3, and OSMI-4, which bind OGT with low-nanomolar affinity. X-ray crystallography showed that the quinolinone-6-sulfonamide scaffold of OSMI compounds act as a uridine mimetic. OSMI-2, OSMI-3, and OSMI-4 have negatively charged carboxylate groups; esterification renders these inhibitors cell-permeable.[28]

Regulation

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Figure 3: Dynamic Competition between glycosylation and phosphorylation of proteins. A: Competition between OGT and kinase for the serine or threonine functional group of a protein. B: Adjacent-site occupancy where O-GlcNAc and O-phosphatase occur next to each other and can influence the turnover or function of proteins reciprocally. The G circle represents an N-acetylglucosamine group, and the P circle represents a phosphate group. Figure adapted from Hart.[29]

O-GlcNAc transferase is part of a dynamic competition for a serine or threonine hydroxyl functional group in a peptide unit. Figure 3 shows an example of both reciprocal same-site occupancy and adjacent-site occupancy. For the same-site occupancy, OGT competes with kinase to catalyze the glycosylation of the protein instead of phosphorylation. The adjacent-site occupancy example shows the naked protein catalyzed by OGT converted to a glycoprotein, which can increase the turnover of proteins such as the tumor repressor p53.[29]

The post-translational modification of proteins by O-GlcNAc is spurred by glucose flux through the hexosamine biosynthetic pathway. OGT catalyzes attachment of the O-GlcNAc group to serine and threonine, while O-GlcNAcase spurs sugar removal.[30][31]

This regulation is important for multiple cellular processes including transcription, signal transduction, and proteasomal degradation. Also, there is competitive regulation between OGT and kinase for the protein to attach to a phosphate group or O-GlcNAc, which can alter the function of proteins in the body through downstream effects.[16][30] OGT inhibits the activity of 6-phosophofructosekinase PFKL by mediating the glycosylation process. This then acts as a part of glycolysis regulation. O-GlcNAc has been defined as a negative transcription regulator in response to steroid hormone signaling.[20]

Studies show that O-GlcNAc transferase interacts directly with the Ten eleven translocation 2 (TET2) enzyme, which converts 5-methylcytosine to 5-hydroxymethylcytosine and regulates gene transcription.[32] Additionally, increasing levels of OGT for O-GlcNAcylation may have therapeutic effects for Alzheimer's disease patients. Brain glucose metabolism is impaired in Alzheimer's disease, and a study suggests that this leads to hyperphosphorylation of tau and degerenation of tau O-GlcNCAcylation. Replenishing tau O-GlcNacylation in the brain along with protein phosphatase could deter this process and improve brain glucose metabolism.[17]

See also

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References

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  1. ^ a b c GRCh38: Ensembl release 89: ENSG00000147162 – Ensembl, May 2017
  2. ^ a b c GRCm38: Ensembl release 89: ENSMUSG00000034160 – Ensembl, May 2017
  3. ^ "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
  4. ^ "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
  5. ^ a b c Lubas WA, Frank DW, Krause M, Hanover JA (April 1997). "O-Linked GlcNAc transferase is a conserved nucleocytoplasmic protein containing tetratricopeptide repeats". The Journal of Biological Chemistry. 272 (14): 9316–24. doi:10.1074/jbc.272.14.9316. PMID 9083068.
  6. ^ a b "Entrez Gene: OGT O-linked N-acetylglucosamine (GlcNAc) transferase (UDP-N-acetylglucosamine:polypeptide-N-acetylglucosaminyl transferase)".
  7. ^ Banerjee S, Robbins PW, Samuelson J (April 2009). "Molecular characterization of nucleocytosolic O-GlcNAc transferases of Giardia lamblia and Cryptosporidium parvum". Glycobiology. 19 (4): 331–6. doi:10.1093/glycob/cwn107. PMC 2733775. PMID 18948359.
  8. ^ Clarke AJ, Hurtado-Guerrero R, Pathak S, Schüttelkopf AW, Borodkin V, Shepherd SM, et al. (October 2008). "Structural insights into mechanism and specificity of O-GlcNAc transferase". The EMBO Journal. 27 (20): 2780–8. doi:10.1038/emboj.2008.186. PMC 2556091. PMID 18818698.
  9. ^ Rao FV, Dorfmueller HC, Villa F, Allwood M, Eggleston IM, van Aalten DM (April 2006). "Structural insights into the mechanism and inhibition of eukaryotic O-GlcNAc hydrolysis". The EMBO Journal. 25 (7): 1569–78. doi:10.1038/sj.emboj.7601026. PMC 1440316. PMID 16541109.
  10. ^ a b Haltiwanger RS, Blomberg MA, Hart GW (May 1992). "Glycosylation of nuclear and cytoplasmic proteins. Purification and characterization of a uridine diphospho-N-acetylglucosamine:polypeptide beta-N-acetylglucosaminyltransferase". The Journal of Biological Chemistry. 267 (13): 9005–13. doi:10.1016/S0021-9258(19)50380-5. PMID 1533623.
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  12. ^ Maynard JC, Burlingame AL, Medzihradszky KF (November 2016). "Cysteine S-linked N-acetylglucosamine (S-GlcNAcylation), A New Post-translational Modification in Mammals". Molecular & Cellular Proteomics. 15 (11): 3405–3411. doi:10.1074/mcp.M116.061549. PMC 5098038. PMID 27558639.
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  15. ^ Whelan SA, Dias WB, Thiruneelakantapillai L, Lane MD, Hart GW (February 2010). "Regulation of insulin receptor substrate 1 (IRS-1)/AKT kinase-mediated insulin signaling by O-Linked beta-N-acetylglucosamine in 3T3-L1 adipocytes". The Journal of Biological Chemistry. 285 (8): 5204–11. doi:10.1074/jbc.M109.077818. PMC 2820748. PMID 20018868.
  16. ^ a b c d "O15294 (OGT1_HUMAN) Reviewed, UniProtKB/Swiss-Prot". UniProt.
  17. ^ a b Liu F, Shi J, Tanimukai H, Gu J, Gu J, Grundke-Iqbal I, et al. (July 2009). "Reduced O-GlcNAcylation links lower brain glucose metabolism and tau pathology in Alzheimer's disease". Brain. 132 (Pt 7): 1820–32. doi:10.1093/brain/awp099. PMC 2702834. PMID 19451179.
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  19. ^ Yang X, Zhang F, Kudlow JE (July 2002). "Recruitment of O-GlcNAc transferase to promoters by corepressor mSin3A: coupling protein O-GlcNAcylation to transcriptional repression". Cell. 110 (1): 69–80. doi:10.1016/S0092-8674(02)00810-3. PMID 12150998.
  20. ^ a b Yang X, Ongusaha PP, Miles PD, Havstad JC, Zhang F, So WV, et al. (February 2008). "Phosphoinositide signalling links O-GlcNAc transferase to insulin resistance". Nature. 451 (7181): 964–9. Bibcode:2008Natur.451..964Y. doi:10.1038/nature06668. PMID 18288188. S2CID 18459576.
  21. ^ Shafi R, Iyer SP, Ellies LG, O'Donnell N, Marek KW, Chui D, et al. (May 2000). "The O-GlcNAc transferase gene resides on the X chromosome and is essential for embryonic stem cell viability and mouse ontogeny". Proceedings of the National Academy of Sciences of the United States of America. 97 (11): 5735–9. Bibcode:2000PNAS...97.5735S. doi:10.1073/pnas.100471497. PMC 18502. PMID 10801981.
  22. ^ Chen Q, Chen Y, Bian C, Fujiki R, Yu X (January 2013). "TET2 promotes histone O-GlcNAcylation during gene transcription". Nature. 493 (7433): 561–4. Bibcode:2013Natur.493..561C. doi:10.1038/nature11742. PMC 3684361. PMID 23222540.
  23. ^ Daou S, Mashtalir N, Hammond-Martel I, Pak H, Yu H, Sui G, et al. (February 2011). "Crosstalk between O-GlcNAcylation and proteolytic cleavage regulates the host cell factor-1 maturation pathway". Proceedings of the National Academy of Sciences of the United States of America. 108 (7): 2747–52. Bibcode:2011PNAS..108.2747D. doi:10.1073/pnas.1013822108. PMC 3041071. PMID 21285374.
  24. ^ Meek RW, Blaza JN, Busmann JA, Alteen MG, Vocadlo DJ, Davies GJ (11 November 2021). "Cryo-EM structure provides insights into the dimer arrangement of the O-linked β-N-acetylglucosamine transferase OGT". Nature Communications. 12 (1): 6508. Bibcode:2021NatCo..12.6508M. doi:10.1038/s41467-021-26796-6. ISSN 2041-1723. PMC 8586251. PMID 34764280.
  25. ^ Cheng SS, Mody AC, Woo CM (2024-11-07). "Opportunities for Therapeutic Modulation of O-GlcNAc". Chemical Reviews. doi:10.1021/acs.chemrev.4c00417. ISSN 0009-2665.
  26. ^ a b Gloster TM, Zandberg WF, Heinonen JE, Shen DL, Deng L, Vocadlo DJ (March 2011). "Hijacking a biosynthetic pathway yields a glycosyltransferase inhibitor within cells". Nature Chemical Biology. 7 (3): 174–81. doi:10.1038/nchembio.520. PMC 3202988. PMID 21258330.
  27. ^ a b Ortiz-Meoz RF, Jiang J, Lazarus MB, Orman M, Janetzko J, Fan C, et al. (June 2015). "A small molecule that inhibits OGT activity in cells". ACS Chemical Biology. 10 (6): 1392–7. doi:10.1021/acschembio.5b00004. PMC 4475500. PMID 25751766.
  28. ^ Martin SE, Tan ZW, Itkonen HM, Duveau DY, Paulo JA, Janetzko J, et al. (October 2018). "Structure-Based Evolution of Low Nanomolar O-GlcNAc Transferase Inhibitors". Journal of the American Chemical Society. 140 (42): 13542–13545. doi:10.1021/jacs.8b07328. PMC 6261342. PMID 30285435.
  29. ^ a b Hart GW, Housley MP, Slawson C (April 2007). "Cycling of O-linked beta-N-acetylglucosamine on nucleocytoplasmic proteins". Nature. 446 (7139): 1017–22. Bibcode:2007Natur.446.1017H. doi:10.1038/nature05815. PMID 17460662. S2CID 4392021.
  30. ^ a b Yang X, Su K, Roos MD, Chang Q, Paterson AJ, Kudlow JE (June 2001). "O-linkage of N-acetylglucosamine to Sp1 activation domain inhibits its transcriptional capability". Proceedings of the National Academy of Sciences of the United States of America. 98 (12): 6611–6. Bibcode:2001PNAS...98.6611Y. doi:10.1073/pnas.111099998. PMC 34401. PMID 11371615.
  31. ^ Love DC, Hanover JA (November 2005). "The hexosamine signaling pathway: deciphering the "O-GlcNAc code"". Science's STKE. 2005 (312): re13. doi:10.1126/stke.3122005re13. PMID 16317114. S2CID 29082840.
  32. ^ Tahiliani M, Koh KP, Shen Y, Pastor WA, Bandukwala H, Brudno Y, et al. (May 2009). "Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1". Science. 324 (5929): 930–5. Bibcode:2009Sci...324..930T. doi:10.1126/science.1170116. PMC 2715015. PMID 19372391.
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  • The O-GlcNAc Database[1][2] - A curated database for protein O-GlcNAcylation and referencing more than 14 000 protein entries and 10 000 O-GlcNAc sites.
 
Suzanne Walker describes The split personality of human O-GlcNAc transferase during a seminar at the US NIH, April 18, 2017
  1. ^ Wulff-Fuentes E, Berendt RR, Massman L, Danner L, Malard F, Vora J, Kahsay R, Olivier-Van Stichelen S (January 2021). "The human O-GlcNAcome database and meta-analysis". Scientific Data. 8 (1): 25. Bibcode:2021NatSD...8...25W. doi:10.1038/s41597-021-00810-4. PMC 7820439. PMID 33479245.
  2. ^ Malard F, Wulff-Fuentes E, Berendt RR, Didier G, Olivier-Van Stichelen S (July 2021). "Automatization and self-maintenance of the O-GlcNAcome catalog: a smart scientific database". Database (Oxford). 2021: 1. doi:10.1093/database/baab039. PMC 8288053. PMID 34279596.
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