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A receptor‐binding protein of C ampylobacter jejuni bacteriophage NCTC 12673 recognizes flagellin glycosylated with acetamidino‐modified pseudaminic acid
Author(s) -
Javed Muhammad Afzal,
Alphen Lieke B.,
Sacher Jessica,
Ding Wen,
Kelly John,
Nargang Cheryl,
Smith David F.,
Cummings Richard D.,
Szymanski Christine M.
Publication year - 2015
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.12849
Subject(s) - flagellin , campylobacter jejuni , biology , microbiology and biotechnology , lytic cycle , bacteriophage , flagellum , campylobacter , mutant , siphoviridae , receptor , biochemistry , bacteria , escherichia coli , virology , gene , genetics , virus
Summary Bacteriophage receptor‐binding proteins ( RBPs ) confer host specificity. We previously identified a putative RBP ( G p047) from the campylobacter lytic phage NCTC 12673 and demonstrated that G p047 has a broader host range than its parent phage. While NCTC 12673 recognizes the capsular polysaccharide ( CPS ) of a limited number of C ampylobacter jejuni isolates, G p047 binds to a majority of C . jejuni and related C ampylobacter coli strains. In this study, we demonstrate that G p047 also binds to acapsular mutants, suggesting that unlike the parent phage, CPS is not the receptor for G p047. Affinity chromatography and far‐western analyses of C . jejuni lysates using G p047 followed by mass spectrometry indicated that G p047 binds to the major flagellin protein, FlaA . Because C . jejuni flagellin is extensively glycosylated, we investigated this binding specificity further and demonstrate that G p047 only recognizes flagellin decorated with acetamidino‐modified pseudaminic acid. This binding activity is localized to the C ‐terminal quarter of the protein and both wild‐type and coccoid forms of C . jejuni are recognized. In addition, G p047 treatment agglutinates vegetative cells and reduces their motility. Because G p047 is highly conserved among all campylobacter phages sequenced to date, it is likely that this protein plays an important role in the phage life cycle.