Myosin ATP turnover rate is a mechanism involved in thermogenesis in resting skeletal muscle fibers.
Thermogenesis by resting muscle varies with conditions and plays an active role in homeostasis of body weight. The low metabolic rate of living resting muscles requires that ATP turnover by myosin be inhibited relative to the purified protein in vitro. This inhibition has not been previously seen in in vitro systems. We used quantitative epifluorescence microscopy of fluorescent nucleotides to measure single nucleotide turnovers in relaxed, permeable skeletal muscle fibers. We observed two lifetimes for nucleotide release by myosin: a fast component with a lifetime of approximately 20 s, similar to that of purified myosin, and a slower component with a lifetime of 230 +/- 24 s. We define the latter component to be the "super relaxed state." The fraction of myosins in the super relaxed state was decreased at lower temperatures, by substituting GTP for ATP or by increased levels of myosin phosphorylation. All of these conditions have also been shown to cause increased disorder in the structure of the thick filament. We propose a model in which the structure of the thick filament modulates the nucleotide turnover rates of myosin in relaxed fibers. Modulation of the relative populations of the super relaxed and conventional relaxed states could have a profound effect on muscle thermogenesis, with the capacity to also significantly alter whole-body metabolic rate.
Stewart
Melanie A
MA
Department of Biochemistry and Biophysics, Cardiovascular Research Institute, University of California, San Francisco, CA 94158, USA.
Franks-Skiba
Kathleen
K
Chen
Susan
S
Cooke
Roger
R
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism*; D000266:Adenylyl Imidodiphosphate / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D004734:Energy Metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D018485:Muscle Fibers, Skeletal / Q000378:metabolism* / Q000648:ultrastructure; D009126:Muscle Relaxation / Q000502:physiology; D009218:Myosins / Q000378:metabolism*; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism; D011817:Rabbits; D022722:Thermogenesis / Q000502:physiology*
C523356:2'(3')-O-methylanthraniloyl-gamma-amido-adenosine triphosphate; D005456:Fluorescent Dyes; D009711:Nucleotides; D000266:Adenylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D009218:Myosins