Localization by site-directed mutagenesis of the site in human complement factor H that binds to Streptococcus pyogenes M protein.
M-protein receptors located on Streptococcus pyogenes cells are known to bind human plasma protein factor H. Human factor H is composed of 20 short consensus repeat (SCR) domains containing approximately 60 amino acids each. Factor H controls the activation of the alternative pathway of complement in plasma. We have scanned the entire human factor H molecule by site-directed deletion mutagenesis, expressed the recombinant proteins in insect cells using the baculovirus system, and measured the binding of different purified mutant proteins to three strains of S. pyogenes. These studies have revealed that recombinant factor H lacking SCR domains 6 to 10 does not bind to wild-type M+ S. pyogenes JRS4. Experiments performed with S. pyogenes JRS251, in which both C-repeat domains of M protein were deleted, demonstrated that all of the factor H mutant proteins bound weakly to these cells except those lacking the SCR region from domains 6 to 10. Neither human factor H nor any of the recombinant proteins bound to the M- strain JRS145. Our results indicate that the only binding site on human factor H that interacts with streptococcus M protein is located in SCR domains 6 to 10 of factor H and that regions of M protein outside the C-repeat domains are involved in binding factor H.
Sharma
A K
AK
Department of Biochemistry, The University of Texas Health Science Center, Tyler 75710-2003, USA.
Pangburn
M K
MK
D000818:Animals; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D002352:Carrier Proteins; D017242:Complement Factor H / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera; D013297:Streptococcus pyogenes / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C493495:CFH protein, human; D002352:Carrier Proteins; D011994:Recombinant Proteins; C014689:streptococcal M protein; D017242:Complement Factor H