Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus

Glycosylation is a widespread mechanism employed by both eukaryotes and bacteria to increase the functional diversity of their proteomes. The nontypeable Haemophilus influenzae glycosyltransferase HMW1C mediates unconventional N-linked glycosylation of the adhesive protein HMW1, which is encoded in...

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Main Authors:	Rempe, Katherine A., Spruce, Lynn A., Porsch, Eric A., Seeholzer, Steven H., Nørskov-Lauritsen, Niels, St. Geme, Joseph W.
Format:	Online
Language:	English
Published:	American Society of Microbiology 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4550697/

id	pubmed-4550697
recordtype	oai_dc
spelling	pubmed-45506972015-08-27 Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus Rempe, Katherine A. Spruce, Lynn A. Porsch, Eric A. Seeholzer, Steven H. Nørskov-Lauritsen, Niels St. Geme, Joseph W. Research Article Glycosylation is a widespread mechanism employed by both eukaryotes and bacteria to increase the functional diversity of their proteomes. The nontypeable Haemophilus influenzae glycosyltransferase HMW1C mediates unconventional N-linked glycosylation of the adhesive protein HMW1, which is encoded in a two-partner secretion system gene cluster that also encodes HMW1C. In this system, HMW1 is modified in the cytoplasm by sequential transfer of hexose residues. In the present study, we examined Kingella kingae and Aggregatibacter aphrophilus homologues of HMW1C that are not encoded near a gene encoding an obvious acceptor protein. We found both homologues to be functional glycosyltransferases and identified their substrates as the K. kingae Knh and the A. aphrophilus EmaA trimeric autotransporter proteins. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed multiple sites of N-linked glycosylation on Knh and EmaA. Without glycosylation, Knh and EmaA failed to facilitate wild-type levels of bacterial autoaggregation or adherence to human epithelial cells, establishing that glycosylation is essential for proper protein function. American Society of Microbiology 2015-08-25 /pmc/articles/PMC4550697/ /pubmed/26307167 http://dx.doi.org/10.1128/mBio.01206-15 Text en Copyright © 2015 Rempe et al. http://creativecommons.org/licenses/by-nc-sa/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license (http://creativecommons.org/licenses/by-nc-sa/3.0/) , which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Rempe, Katherine A. Spruce, Lynn A. Porsch, Eric A. Seeholzer, Steven H. Nørskov-Lauritsen, Niels St. Geme, Joseph W.
spellingShingle	Rempe, Katherine A. Spruce, Lynn A. Porsch, Eric A. Seeholzer, Steven H. Nørskov-Lauritsen, Niels St. Geme, Joseph W. Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus
author_facet	Rempe, Katherine A. Spruce, Lynn A. Porsch, Eric A. Seeholzer, Steven H. Nørskov-Lauritsen, Niels St. Geme, Joseph W.
author_sort	Rempe, Katherine A.
title	Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus
title_short	Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus
title_full	Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus
title_fullStr	Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus
title_full_unstemmed	Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus
title_sort	unconventional n-linked glycosylation promotes trimeric autotransporter function in kingella kingae and aggregatibacter aphrophilus
description	Glycosylation is a widespread mechanism employed by both eukaryotes and bacteria to increase the functional diversity of their proteomes. The nontypeable Haemophilus influenzae glycosyltransferase HMW1C mediates unconventional N-linked glycosylation of the adhesive protein HMW1, which is encoded in a two-partner secretion system gene cluster that also encodes HMW1C. In this system, HMW1 is modified in the cytoplasm by sequential transfer of hexose residues. In the present study, we examined Kingella kingae and Aggregatibacter aphrophilus homologues of HMW1C that are not encoded near a gene encoding an obvious acceptor protein. We found both homologues to be functional glycosyltransferases and identified their substrates as the K. kingae Knh and the A. aphrophilus EmaA trimeric autotransporter proteins. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed multiple sites of N-linked glycosylation on Knh and EmaA. Without glycosylation, Knh and EmaA failed to facilitate wild-type levels of bacterial autoaggregation or adherence to human epithelial cells, establishing that glycosylation is essential for proper protein function.
publisher	American Society of Microbiology
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4550697/
_version_	1613469455236464640

Unconventional N-Linked Glycosylation Promotes Trimeric Autotransporter Function in Kingella kingae and Aggregatibacter aphrophilus

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