Skeletal muscle myosin promotes coagulation by binding factor XI via its A3 domain and enhancing thrombin-induced factor XI activation

• Published in: The Journal of biological chemistry Vol. 298; no. 2; p. 101567
• Main Authors: Morla, Shravan, Deguchi, Hiroshi, Zilberman-Rudenko, Jevgenia, Gruber, András, McCarty, Owen J.T., Srivastava, Priyanka, Gailani, David, Griffin, John H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-02-2022; American Society for Biochemistry and Molecular Biology
• Subjects: Accelerated Communication; Antibodies, Monoclonal - chemistry; Binding Sites; Blood Coagulation; coagulation factor; factor XI; Factor XI - chemistry; Factor XI - genetics; Factor XI - metabolism; myosin; Prekallikrein - chemistry; Prekallikrein - metabolism; Protein Domains; Recombinant Fusion Proteins - chemistry; skeletal muscle myosin; Skeletal Muscle Myosins - metabolism; thrombin; Thrombin - metabolism; A; thrombin; factor XI; PS; ALP; myosin; mAb; BSA; rFXI; TF; SkM; FXI; skeletal muscle myosin; coagulation factor; PK; BLI; PolyP
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/j.jbc.2022.101567

Skeletal muscle myosin (SkM) has been shown to possess procoagulant activity; however, the mechanisms of this coagulation-enhancing activity involving plasma coagulation pathways and factors are incompletely understood. Here, we discovered direct interactions between immobilized SkM and coagulation factor XI (FXI) using biolayer interferometry (Kd = 0.2 nM). In contrast, we show that prekallikrein, a FXI homolog, did not bind to SkM, reflecting the specificity of SkM for FXI binding. We also found that the anti-FXI monoclonal antibody, mAb 1A6, which recognizes the Apple (A) 3 domain of FXI, potently inhibited binding of FXI to immobilized SkM, implying that SkM binds FXI A3 domain. In addition, we show that SkM enhanced FXI activation by thrombin in a concentration-dependent manner. We further used recombinant FXI chimeric proteins in which each of the four A domains of the heavy chain (designated A1 through A4) was individually replaced with the corresponding A domain from prekallikrein to investigate SkM-mediated enhancement of thrombin-induced FXI activation. These results indicated that activation of two FXI chimeras with substitutions of either the A3 domains or A4 domains was not enhanced by SkM, whereas substitution of the A2 domain did not reduce the thrombin-induced activation compared with wildtype FXI. These data strongly suggest that functional interaction sites on FXI for SkM involve the A3 and A4 domains. Thus, this study is the first to reveal and support the novel intrinsic blood coagulation pathway concept that the procoagulant mechanisms of SkM include FXI binding and enhancement of FXI activation by thrombin.