Solution structural model of the complex of the binding regions of human plasminogen with its M-protein receptor from Streptococcus pyogenes

Abstract

VEK50 is a truncated peptide from a Streptococcal pyogenes surface human plasminogen (hPg) binding M-protein (PAM). VEK50 contains the full A-domain of PAM, which is responsible for its low nanomolar binding to hPg. The interaction of VEK50 with kringle 2, the PAM-binding domain in hPg (K2_hPg), has been studied by high-resolution NMR spectroscopy. The data show that each VEK50 monomer in solution contains two tight binding sites for K2_hPg, one each in the a1- (RH1; R¹⁷H¹⁸) and a2- (RH2; R³⁰H³¹) repeats within the A-domain of VEK50. Two mutant forms of VEK50, viz., VEK50[RH1/AA] (VEK50^ΔRH1) and VEK50[RH2/AA] (VEK50^ΔRH2), were designed by replacing each RH with AA, thus eliminating one of the K2_hPg binding sites within VEK50, and allowing separate study of each binding site. Using ¹³C- and ¹⁵N-labeled peptides, NMR-derived solution structures of VEK50 in its complex with K2_hPg were solved. We conclude that the A-domain of PAM can accommodate two molecules of K2_hPg docked within a short distance of each other, and the strength of the binding is slightly different for each site. The solution structure of the VEK50/K2_hPg, complex, which is a reductionist model of the PAM/hPg complex, provides insights for the binding mechanism of PAM to a host protein, a process that is critical to S. pyogenes virulence.

Keywords: 3-D solution structure; A-repeats; Bacterial receptor; Modular proteins; NMR structures; Peptide mutagenesis; Plasminogen binding.