Homologous mutations in human β, embryonic, and perinatal muscle myosins have divergent effects on molecular power generation

• 成果类型: Article
• 署名作者: Liu, Chao; Karabina, Anastasia; Meller, Artur; Bhattacharjee, Ayan; Agostino, Colby J.; Bowman, Greg R.; Ruppel, Kathleen M.; Spudich, James A.; Leinwand, Leslie A.
• 署名单位: Stanford University; Stanford University; United States Department of Energy (DOE); Lawrence Livermore National Laboratory; University of Colorado System; University of Colorado Boulder; University of Colorado System; University of Colorado Boulder; Washington University (WUSTL); Washington University (WUSTL); University of Pennsylvania
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14942
• DOI: 10.1073/pnas.2315472121
• 发表日期: 2024-02-27
• 关键词: load-dependent kinetics; trismus-pseudocamptodactyly syndrome; hypertrophic cardiomyopathy; cardiac myosin; sheldon-syndrome; r403q mutation; skeletal; FORCE; velocity; actin

摘要

Mutations at a highly conserved homologous residue in three closely related muscle myosins cause three distinct diseases involving muscle defects: R671C in beta- cardiac myosin causes hypertrophic cardiomyopathy, R672C and R672H in embryonic skeletal myosin cause Freeman-Sheldon syndrome, and R674Q in perinatal skeletal myosin causes trismus- pseudocamptodactyly syndrome. It is not known whether their effects at the molecular level are similar to one another or correlate with disease phenotype and severity. To this end, we investigated the effects of the homologous mutations on key factors of molecular power production using recombinantly expressed human beta, embryonic, and perinatal myosin subfragment- 1. We found large effects in the developmental myosins but minimal effects in beta myosin, and magnitude of changes correlated partially with clinical severity. The mutations in the developmental myosins dramatically decreased the step size and load- sensitive actin- detachment rate of single molecules measured by optical tweezers, in addition to decreasing overall enzymatic (ATPase) cycle rate. In contrast, the only measured effect of R671C in beta myosin was a larger step size. Our measurements of step size and bound times predicted velocities consistent with those measured in an in vitro motility assay. Finally, molecular dynamics simulations predicted that the arginine to cysteine mutation in embryonic, but not beta, myosin may reduce pre- powerstroke lever arm priming and ADP pocket opening, providing a possible structural mechanism consistent with the experimental observations. This paper presents direct comparisons of homologous mutations in several different myosin isoforms, whose divergent functional effects are a testament to myosin's highly allosteric nature.