Nucleotide specificity of canine cardiac sarcoplasmic reticulum. Differential alteration of enzyme properties by detergent treatment.
We previously demonstrated that, in contrast to the hydrolysis of ATP, the hydrolysis of GTP by canine cardiac sarcoplasmic reticulum is not sensitive to calcium. Based on a variety of qualitative and quantitative considerations (cf. Tate, C. A., Bick, R. J., Chu, A., Van Winkle, W. B., and Entman, M. L. (1985) J. Biol. Chem. 260, 9618-9623), we suggested that the hydrolysis of ATP and GTP appears to be effected by the same enzyme. In the present paper, we examined the sensitivity of both enzymatic activities to low concentrations of detergent. With nonsolubilizing concentrations of the nonionic detergent, octaethylene glycol monododecyl ether, the hydrolysis of GTP was rendered partially calcium-sensitive resulting from a slightly increased total (Ca2+ + Mg2+)-GTPase activity and a markedly inhibited calcium-independent (Mg2+-dependent) GTPase activity. Calcium-dependent ATPase activity was increased with octaethylene glycol monododecyl ether, mimicking the effect of the ionophore, A23187. Calcium-dependent ATPase activity and detergent-induced calcium-dependent GTPase activity were similar in (a) calcium sensitivity, (b) sensitivity to mersalyl, and (c) pressure inactivation through dilution and centrifugation, all of which differed from the untreated calcium-independent GTPase activity. Calcium-dependent ATPase activity differed from calcium-dependent GTPase activity with (a) a higher nucleotide affinity, (b) a lower vanadate sensitivity, and (c) a calcium sensitivity for phosphoenzyme formation. Thus, the detergent-induced perturbation of the GTPase resulted in an enzyme with many characteristics qualitatively and quantitatively similar to the calcium ATPase.
Tate
C A
CA
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.
Bick
R J
RJ
Blaylock
S L
SL
Youker
K A
KA
Scherer
N M
NM
Entman
M L
ML
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D004285:Dogs; D004789:Enzyme Activation; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008634:Mersalyl / Q000494:pharmacology; D009206:Myocardium / Q000201:enzymology*; D009711:Nucleotides / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D011312:Pressure; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology*; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000494:pharmacology*; D014638:Vanadates / Q000494:pharmacology
D003902:Detergents; D009711:Nucleotides; D013501:Surface-Active Agents; C017272:dodecyloctaethyleneglycol monoether; D000001:Calcimycin; D014638:Vanadates; D011092:Polyethylene Glycols; D008634:Mersalyl; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D020558:GTP Phosphohydrolases; D000252:Calcium-Transporting ATPases; D008274:Magnesium; D002118:Calcium