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 (K2hPg), has been studied by high-resolution NMR spectroscopy. The data show that each VEK50 monomer in solution contains two tight binding sites for K2hPg, 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] (VEK50ΔRH1) and VEK50[RH2/AA] (VEK50ΔRH2), were designed by replacing each RH with AA, thus eliminating one of the K2hPg 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 K2hPg were solved. We conclude that the A-domain of PAM can accommodate two molecules of K2hPg 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/K2hPg, 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.
| Original language | English |
|---|---|
| Pages (from-to) | 18-29 |
| Number of pages | 12 |
| Journal | Journal of Structural Biology |
| Volume | 208 |
| DOIs | |
| Publication status | Published - 1 Oct 2019 |
Keywords
- 3-D solution structure
- A-repeats
- Bacterial receptor
- Modular proteins
- NMR structures
- Peptide mutagenesis
- Plasminogen binding
Cite this
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Solution structural model of the complex of the binding regions of human plasminogen with its M-protein receptor from Streptococcus pyogenes. / Yuan, Yue; Ayinuola, Yetunde A.; Singh, Damini; Ayinuola, Olawole; Mayfield, Jeffrey A.; Quek, Adam; Whisstock, James C.; Law, Ruby H.P.; Lee, Shaun W.; Ploplis, Victoria A.; Castellino, Francis J.
In: Journal of Structural Biology, Vol. 208, 01.10.2019, p. 18-29.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Solution structural model of the complex of the binding regions of human plasminogen with its M-protein receptor from Streptococcus pyogenes
AU - Yuan, Yue
AU - Ayinuola, Yetunde A.
AU - Singh, Damini
AU - Ayinuola, Olawole
AU - Mayfield, Jeffrey A.
AU - Quek, Adam
AU - Whisstock, James C.
AU - Law, Ruby H.P.
AU - Lee, Shaun W.
AU - Ploplis, Victoria A.
AU - Castellino, Francis J.
PY - 2019/10/1
Y1 - 2019/10/1
N2 - 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 (K2hPg), has been studied by high-resolution NMR spectroscopy. The data show that each VEK50 monomer in solution contains two tight binding sites for K2hPg, 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] (VEK50ΔRH1) and VEK50[RH2/AA] (VEK50ΔRH2), were designed by replacing each RH with AA, thus eliminating one of the K2hPg 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 K2hPg were solved. We conclude that the A-domain of PAM can accommodate two molecules of K2hPg 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/K2hPg, 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.
AB - 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 (K2hPg), has been studied by high-resolution NMR spectroscopy. The data show that each VEK50 monomer in solution contains two tight binding sites for K2hPg, 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] (VEK50ΔRH1) and VEK50[RH2/AA] (VEK50ΔRH2), were designed by replacing each RH with AA, thus eliminating one of the K2hPg 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 K2hPg were solved. We conclude that the A-domain of PAM can accommodate two molecules of K2hPg 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/K2hPg, 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.
KW - 3-D solution structure
KW - A-repeats
KW - Bacterial receptor
KW - Modular proteins
KW - NMR structures
KW - Peptide mutagenesis
KW - Plasminogen binding
UR - http://www.scopus.com/inward/record.url?scp=85068841670&partnerID=8YFLogxK
U2 - 10.1016/j.jsb.2019.07.005
DO - 10.1016/j.jsb.2019.07.005
M3 - Article
VL - 208
SP - 18
EP - 29
JO - Journal of Structural Biology
JF - Journal of Structural Biology
SN - 1047-8477
ER -