Identification of residues within ligand-binding domain 1 (LBD1) of the Borrelia burgdorferi OspC protein required for function in the mammalian environment.
Borrelia burgdorferi outer surface protein C (ospC) is required for the establishment of infection in mammals. However, its precise function remains controversial. The biologically active form of OspC appears to be a homodimer. Alpha helix 1 and 1' of the apposing monomers form a solvent-accessible pocket at the dimeric interface that presents a putative ligand-binding domain (LBD1). Here we employ site-directed and allelic-exchange mutagenesis to test the hypothesis that LBD1 is a determinant of OspC function in the mammalian environment. Substitution of residues K60, E61 and E63 which line LBD1 resulted in the loss of infectivity or influenced dissemination. Analyses of the corresponding recombinant proteins demonstrated that the loss of function was not due to structural perturbation, impaired dimer formation or the loss of plasminogen binding. This study is the first to assess the involvement of individual residues and domains of OspC in its in vivo function. The data support the hypothesis that OspC interacts with a mammalian derived ligand that is critical for survival during early infection. These results shed new light on the structure-functions relationships of OspC and challenge existing hypotheses regarding OspC function in mammals.
Earnhart
Christopher G
CG
Department of Microbiology and Immunology, Medical College of Virginia at Virginia Commonwealth University, Richmond, VA, USA.
Leblanc
Delacy V
DV
Alix
Katie E
KE
Desrosiers
Daniel C
DC
Radolf
Justin D
JD
Marconi
Richard T
RT
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D025065:Borrelia burgdorferi / Q000235:genetics / Q000472:pathogenicity*; D019281:Dimerization; D055786:Gene Knockout Techniques; D051379:Mice; D008809:Mice, Inbred C3H; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D050505:Mutant Proteins; C079616:OspC protein; D037521:Virulence Factors; D010958:Plasminogen