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New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein

Serine-rich repeat glycoproteins (SRRPs) conserved in streptococci and staphylococci are important for bacterial colonization and pathogenesis. Fap1, a well studied SRRP is a major surface constituent of Streptococcus parasanguinis and is required for bacterial adhesion and biofilm formation. Biogen...

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Detalles Bibliográficos
Autores principales: Zhang, Hua, Zhou, Meixian, Yang, Tiandi, Haslam, Stuart M., Dell, Anne, Wu, Hui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5063993/
https://www.ncbi.nlm.nih.gov/pubmed/27539847
http://dx.doi.org/10.1074/jbc.M116.731695
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author Zhang, Hua
Zhou, Meixian
Yang, Tiandi
Haslam, Stuart M.
Dell, Anne
Wu, Hui
author_facet Zhang, Hua
Zhou, Meixian
Yang, Tiandi
Haslam, Stuart M.
Dell, Anne
Wu, Hui
author_sort Zhang, Hua
collection PubMed
description Serine-rich repeat glycoproteins (SRRPs) conserved in streptococci and staphylococci are important for bacterial colonization and pathogenesis. Fap1, a well studied SRRP is a major surface constituent of Streptococcus parasanguinis and is required for bacterial adhesion and biofilm formation. Biogenesis of Fap1 is a multistep process that involves both glycosylation and secretion. A series of glycosyltransferases catalyze sequential glycosylation of Fap1. We have identified a unique hybrid protein dGT1 (dual glycosyltransferase 1) that contains two distinct domains. N-terminal DUF1792 is a novel GT-D-type glycosyltransferase, transferring Glc residues to Glc-GlcNAc-modified Fap1. C-terminal dGT1 (CgT) is predicted to possess a typical GT-A-type glycosyltransferase, however, the activity remains unknown. In this study, we determine that CgT is a distinct glycosyltransferase, transferring GlcNAc residues to Glc-Glc-GlcNAc-modified Fap1. A 2.4-Å x-ray crystal structure reveals that CgT has a unique binding domain consisting of three α helices in addition to a typical GT-A-type glycosyltransferase domain. The helical domain is crucial for the oligomerization of CgT. Structural and biochemical studies revealed that the helix domain is required for the protein-protein interaction and crucial for the glycosyltransferase activity of CgT in vitro and in vivo. As the helix domain presents a novel structural fold, we conclude that CgT represents a new member of GT-A-type glycosyltransferases.
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spelling pubmed-50639932016-10-26 New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein Zhang, Hua Zhou, Meixian Yang, Tiandi Haslam, Stuart M. Dell, Anne Wu, Hui J Biol Chem Microbiology Serine-rich repeat glycoproteins (SRRPs) conserved in streptococci and staphylococci are important for bacterial colonization and pathogenesis. Fap1, a well studied SRRP is a major surface constituent of Streptococcus parasanguinis and is required for bacterial adhesion and biofilm formation. Biogenesis of Fap1 is a multistep process that involves both glycosylation and secretion. A series of glycosyltransferases catalyze sequential glycosylation of Fap1. We have identified a unique hybrid protein dGT1 (dual glycosyltransferase 1) that contains two distinct domains. N-terminal DUF1792 is a novel GT-D-type glycosyltransferase, transferring Glc residues to Glc-GlcNAc-modified Fap1. C-terminal dGT1 (CgT) is predicted to possess a typical GT-A-type glycosyltransferase, however, the activity remains unknown. In this study, we determine that CgT is a distinct glycosyltransferase, transferring GlcNAc residues to Glc-Glc-GlcNAc-modified Fap1. A 2.4-Å x-ray crystal structure reveals that CgT has a unique binding domain consisting of three α helices in addition to a typical GT-A-type glycosyltransferase domain. The helical domain is crucial for the oligomerization of CgT. Structural and biochemical studies revealed that the helix domain is required for the protein-protein interaction and crucial for the glycosyltransferase activity of CgT in vitro and in vivo. As the helix domain presents a novel structural fold, we conclude that CgT represents a new member of GT-A-type glycosyltransferases. American Society for Biochemistry and Molecular Biology 2016-10-14 2016-08-17 /pmc/articles/PMC5063993/ /pubmed/27539847 http://dx.doi.org/10.1074/jbc.M116.731695 Text en © 2016 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version free via Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) .
spellingShingle Microbiology
Zhang, Hua
Zhou, Meixian
Yang, Tiandi
Haslam, Stuart M.
Dell, Anne
Wu, Hui
New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein
title New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein
title_full New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein
title_fullStr New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein
title_full_unstemmed New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein
title_short New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein
title_sort new helical binding domain mediates a glycosyltransferase activity of a bifunctional protein
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5063993/
https://www.ncbi.nlm.nih.gov/pubmed/27539847
http://dx.doi.org/10.1074/jbc.M116.731695
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