Identification of the IgA-binding region in streptococcal protein Arp.
Cell surface proteins that bind to the Fc part of human IgA are expressed by different species of pathogenic streptococci. The most extensively characterized streptococcal IgA-binding protein is the Streptococcus pyogenes protein Arp4, a member of the M protein family. Here we describe work that identifies the IgA-binding region in this streptococcal protein. A comparison of the amino acid sequences of protein Arp4 and four other IgA-binding proteins of S. pyogenes first made possible the identification of a putative IgA-binding region. Site-specific mutagenesis and generation of deletions were then used to show that Arp4 derivatives lacking different parts of the putative IgA-binding region had lost the ability to bind IgA. Conclusive evidence for the localization of the IgA-binding region was obtained through the characterization of a chimeric protein, in which the putative IgA-binding region of Arp4 had been introduced into another S. pyogenes cell surface protein that does not bind IgA. Our data show that a region comprising 29-amino acid residues in the N-terminal part of Arp4 is necessary and sufficient for IgA-binding capacity. Competitive inhibition experiments with synthetic peptides indicated that the C-terminal half of this 29 residue region may be most important for the IgA-binding property of Arp4. These results identify, for the first time, the ligand-binding region in an Fc alpha binding protein.
Johnsson
E
E
Department of Microbiology, Lund University, Sweden.
Andersson
G
G
Lindahl
G
G
Hedén
L O
LO
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000737:chemistry*; D007070:Immunoglobulin A / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013297:Streptococcus pyogenes / Q000737:chemistry; D013329:Structure-Activity Relationship
C062462:Arp4 protein, Streptococcus pyogenes; D001426:Bacterial Proteins; D002352:Carrier Proteins; D007070:Immunoglobulin A; D010455:Peptides; D011993:Recombinant Fusion Proteins