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Title: Unraveling the sequence of cytosolic reactions in the export of GspB adhesin from Streptococcus gordonii

Journal Article · · Journal of Biological Chemistry
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Many pathogenic bacteria, including Streptococcus gordonii, possess a pathway for the cellular export of a single serine-rich-repeat protein that mediates the adhesion of bacteria to host cells and the extracellular matrix. This adhesin protein is O-glycosylated by several cytosolic glycosyltransferases and requires three accessory Sec proteins (Asp1–3) for export, but how the adhesin protein is processed for export is not well understood. Here, in this paper, we report that the S. gordonii adhesin GspB is sequentially O-glycosylated by three enzymes (GtfA/B, Nss, and Gly) that attach N-acetylglucosamine and glucose to Ser/Thr residues. We also found that modified GspB is transferred from the last glycosyltransferase to the Asp1/2/3 complex. Crystal structures revealed that both Asp1 and Asp3 are related to carbohydrate-binding proteins, suggesting that they interact with carbohydrates and bind glycosylated adhesin, a notion that was supported by further analyses. We further observed that Asp1 also has an affinity for phospholipids, which is attenuated by Asp2. In summary, our findings support a model in which the GspB adhesin is sequentially glycosylated by GtfA/B, Nss, and Gly and then transferred to the Asp1/2/3 complex in which Asp1 mediates the interaction of the Asp1/2/3 complex with the lipid bilayer for targeting of matured GspB to the export machinery.

Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
National Institutes of Health (NIH); Howard Hughes Medical Institute–Helen Hay Whitney Foundation; US Department of Veterans Affairs; Northern California Institute for Research and Education; USDOE Office of Science (SC)
Grant/Contract Number:
SC0015662; GM052586; 1S10OD018530; P41GM10349010; R01-AI41513; R01-AI106987; P41 GM103403; S10 RR029205; AC02-06CH11357
OSTI ID:
1769426
Alternate ID(s):
OSTI ID: 1434746
Journal Information:
Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Vol. 293 Journal Issue: 14; ISSN 0021-9258
Publisher:
ElsevierCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 10 works
Citation information provided by
Web of Science

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Cited By (2)

Membrane trafficking of the bacterial adhesin GspB and the accessory Sec transport machinery journal December 2018
Serine-rich repeat proteins from gut microbes journal April 2019

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
bacterial pathogenesis
glycosylation
adhesin
crystallography
protein export
protein secretion
glycosyltransferase