Comparison of sarcoplasmic reticulum Ca2+-adenosine triphosphatase from vitamin E-deficient dystrophic rabbit skeletal muscle with iron-ascorbate-treated and untreated enzyme.
Sarcoplasmic reticulum Ca2+-ATPase from rabbit skeletal muscle has an Arrhenius curve of enzyme activity with a discontinuity at about 20 degrees C. Preparations treated with FeSO4 and ascorbic acid and from a vitamin E-deficient dystrophic rabbit have 22% of the normal activity and a linear Arrhenius curve (Promkhatkaew, D., Komaratat, P., & Wilairat, P. (1985) Biochem. Int. 10, 937-943). All three preparations were cross-linked to the same extent by dimethyl suberimidate and copper-phenanthroline reagent at temperatures above and below the temperature of the Arrhenius discontinuity. Both iron-ascorbate-treated Ca2+-ATPase and that from a vitamin E-deficient animal had 50% of the normal sulfhydryl content, but the disulfide and free amino contents were unaltered. These observations suggest that loss of sulfhydryl groups through lipid peroxidation, both in vivo and in vitro, resulted in reduction of Ca2+-ATPase activity and loss of the break in the Arrhenius plot. Changes in Ca2+-ATPase polypeptide aggregational state could not account for the discontinuity in the Arrhenius curve as revealed by the similar extent of cross-linking of the three enzyme preparations at temperatures above and below the temperature of the Arrhenius discontinuity.
Nirdnoy
W
W
Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand.
Komaratat
P
P
Wilairat
P
P
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001205:Ascorbic Acid / Q000627:therapeutic use; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D003432:Cross-Linking Reagents; D004220:Disulfides / Q000032:analysis; D007501:Iron / Q000627:therapeutic use; D009137:Muscular Dystrophy, Animal / Q000201:enzymology*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000032:analysis; D014811:Vitamin E Deficiency / Q000201:enzymology*
D000596:Amino Acids; D003432:Cross-Linking Reagents; D004220:Disulfides; D013438:Sulfhydryl Compounds; D007501:Iron; D000252:Calcium-Transporting ATPases; D001205:Ascorbic Acid