The Scl1 protein of M6-type group A Streptococcus binds the human complement regulatory protein, factor H, and inhibits the alternative pathway of complement.
Non-specific activation of the complement system is regulated by the plasma glycoprotein factor H (FH). Bacteria can avoid complement-mediated opsonization and phagocytosis through acquiring FH to the cell surface. Here, we characterize an interaction between the streptococcal collagen-like protein Scl1.6 of M6-type group A Streptococcus (GAS) and FH. Using affinity chromatography with immobilized recombinant Scl1.6 protein, we co-eluted human plasma proteins with molecular weight of 155 kDa, 43 kDa and 38 kDa. Mass spectrometry identified the 155 kDa band as FH and two other bands as isoforms of the FH-related protein-1. The identities of all three bands were confirmed by Western immunoblotting with specific antibodies. Structure-function relation studies determined that the globular domain of the Scl1.6 variant specifically binds FH while fused to collagenous tails of various lengths. This binding is not restricted to Scl1.6 as the phylogenetically linked Scl1.55 variant also binds FH. Functional analyses demonstrated the cofactor activity of the rScl1.6-bound FH for factor I-mediated cleavage of C3b. Finally, purified FH bound to the Scl1.6 protein present in the cell wall material obtained from M6-type GAS. In conclusion, we have identified a functional interaction between Scl1 and plasma FH, which may contribute to GAS evasion of complement-mediated opsonization and phagocytosis.
Caswell
Clayton C
CC
Department of Microbiology, Immunology, and Cell Biology, School of Medicine, West Virginia University, Morgantown, WV 26506, USA.
Han
Runlin
R
Hovis
Kelley M
KM
Ciborowski
Pawel
P
Keene
Douglas R
DR
Marconi
Richard T
RT
Lukomski
Slawomir
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003094:Collagen / Q000235:genetics* / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism; D017242:Complement Factor H / Q000378:metabolism*; D003170:Complement Pathway, Alternative; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010802:Phylogeny; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013297:Streptococcus pyogenes / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C423454:Scl1 protein, Streptococcus; D003179:Complement C3b; D017242:Complement Factor H; D003094:Collagen