Human cardiac myosin- binding protein C restricts actin structural dynamics in a cooperative and phosphorylation-sensitive manner

Thomas A. Bunch, Rhye Samuel Kanassatega, Victoria C. Lepak, Brett A. Colson

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Cardiac myosin- binding protein C (cMyBP-C) is a thick filament-associated protein that influences actin-myosin interactions. cMyBP-C alters myofilament structure and contractile properties in a protein kinase A (PKA) phosphorylation-dependent manner. To determine the effects of cMyBP-C and its phosphorylation on the microsecond rotational dynamics of actin filaments, we attached a phosphorescent probe to F-actin at Cys-374 and performed transient phosphorescence anisotropy (TPA) experiments. Binding of cMyBP-C N-terminal domains (C0 -C2) to labeled F-actin reduced rotational flexibility by 20 -25°, indicated by increased final anisotropy of the TPA decay. The effects of C0 -C2 on actin TPA were highly cooperative (n = -8), suggesting that the cMyBP-C N terminus impacts the rotational dynamics of actin spanning seven monomers (i.e. the length of tropomyosin). PKA-mediated phosphorylation of C0 -C2 eliminated the cooperative effects on actin flexibility and modestly increased actin rotational rates. Effects of Ser to Asp phosphomimetic substitutions in the M-domain of C0 -C2 on actin dynamics only partially recapitulated the phosphorylation effects. C0 -C1 (lacking M-domain/C2) similarly exhibited reduced cooperativity, but not as reduced as by phosphorylated C0 -C2. These results suggest an important regulatory role of the M-domain in cMyBP-C effects on actin structural dynamics. In contrast, phosphomimetic substitution of the glycogen synthase kinase (GSK3β) site in the Pro/Ala-rich linker of C0 -C2 did not significantly affect the TPA results. We conclude that cMyBP-C binding and PKA-mediated phosphorylation can modulate actin dynamics. We propose that these N-terminal cMyBP-C-induced changes in actin dynamics help explain the functional effects of cMyBP-C phosphorylation on actin-myosin interactions.

Original languageEnglish (US)
Pages (from-to)16228-16240
Number of pages13
JournalJournal of Biological Chemistry
Issue number44
StatePublished - Nov 1 2019

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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