Cargando…
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...
Autores principales: | , , , , , |
---|---|
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 |
_version_ | 1782460067361062912 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5063993 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT zhanghua newhelicalbindingdomainmediatesaglycosyltransferaseactivityofabifunctionalprotein AT zhoumeixian newhelicalbindingdomainmediatesaglycosyltransferaseactivityofabifunctionalprotein AT yangtiandi newhelicalbindingdomainmediatesaglycosyltransferaseactivityofabifunctionalprotein AT haslamstuartm newhelicalbindingdomainmediatesaglycosyltransferaseactivityofabifunctionalprotein AT dellanne newhelicalbindingdomainmediatesaglycosyltransferaseactivityofabifunctionalprotein AT wuhui newhelicalbindingdomainmediatesaglycosyltransferaseactivityofabifunctionalprotein |