title: Effects of reduced glycogen on structure and in vitro function of rat sarcoplasmic reticulum Ca2+-ATPase.
abstract: The aim of this study was to examine the effects of reduced glycogen concentration on sarcoplasmic reticulum (SR) Ca(2+)-ATPase activity in rat fast-twitch muscles. In the first experiment, the gastrocnemius (GAS) muscle from one leg was removed, followed by starvation for 24-72 h, after which the remaining GAS was removed. Intra-animal comparisons revealed that starvation caused a 25% reduction (P<0.05) in the glycogen concentration but no change in SR Ca(2+)-ATPase activity in the GAS. In the second experiment, the SR was purified from a mixture of the GAS and vastus lateralis muscles. In half of the samples obtained from each animal, glycogen was extracted from the SR by treatment with glucoamylase. Treatment resulted in a 94.1 and 70.2% decrease (P<0.01) in glycogen and glycogen phosphorylase, respectively, and a 41.5% increase (P<0.05) in a fluorescein isothiocyanate (FITC) binding to SR Ca(2+)-ATPase. On the other hand, SR Ca(2+)-ATPase activity and the affinity of the enzyme for ATP were unaltered. These results do not implicate depletion of muscle glycogen as a contributor to impaired SR Ca(2+)-ATPase activity as measured in vitro. Therefore, it is concluded that muscle glycogen does not influence exercise tolerance and work productivity in working muscles by modulating the structure of protein involved in Ca(2+) sequestering. Furthermore, it is suggested that the FITC binding assay may be inappropriate as a method for examining the mechanisms for the altered activity of SR Ca(2+)-ATPase.
authors.affiliation: Graduate School of Biosphere Science, Hiroshima University, Higashihiroshima-shi, 739-8521, Hiroshima, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Mishima
authors.forename: Minako
authors.initials: M
authors.lastname: Sugiyama
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamada
authors.forename: Makoto
authors.initials: M
authors.lastname: Sakamoto
authors.forename: Masanobu
authors.initials: M
authors.lastname: Wada
mesh_terms: D000818:Animals; D005087:Glucan 1,4-alpha-Glucosidase; D006003:Glycogen / Q000378:metabolism*; D024981:Glycogen Phosphorylase / Q000378:metabolism; D008297:Male; D009132:Muscles / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*; D013217:Starvation / Q000378:metabolism*
chemical_list: D006003:Glycogen; D024981:Glycogen Phosphorylase; D005087:Glucan 1,4-alpha-Glucosidase; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases