Open AccessActin nitrosylation and its effect on myosin driven motility
Author(s) -
Heather M. Bansbach,
William H. Guilford
Publication year - 2016
Publication title -
aims molecular science
Language(s) - English
Resource type - Journals
eISSN - 2372-028X
pISSN - 2372-0301
DOI - 10.3934/molsci.2016.3.426
Subject(s) - heavy meromyosin , myosin , actin , skeletal muscle , chemistry , biophysics , microbiology and biotechnology , protein filament , actin remodeling , s nitrosylation , cytoskeleton , biochemistry , actin cytoskeleton , biology , cysteine , anatomy , cell , enzyme
The cytoskeletal protein actin can be nitrosylated, and others have shown that nitrosylation of actin can affect actin filament polymerization. However, the effects of nitrosylation on its interactions with the motor protein myosin are unknown. We therefore measured the effect of S-nitrosylation on the interactions of several actin isoforms with myosin. We used a modified coumarin switch assay to determine the number of nitrosylated cysteines in α-skeletal muscle, α-smooth muscle, and non-muscle (β and γ) actin in response to in vitro treatment with nitroso-L-cysteine—an endogenous nitric oxide (NO) donor. We also measured actin filament velocity over heavy meromyosin (HMM) using an in vitro motility assay, the isometric force generated by HMM using a laser trap, and the actin activated ATPase rates of HMM. We found that all three isoforms of actin were nitrosylated equally at ~2 sites per monomer. Nitrosylation of skeletal muscle a-actin reduced the velocity of actin filaments over HMM in a dose dependent fashion. The sliding velocities of all actin isoforms over HMM were reduced equally by ~24% when nitrosylated with 50 µM donor. Our data are consistent with actin nitrosylation causing an increase in the time myosin remains bound to actin during its hydrolytic cycle