Detergent activation of the ATPase activity of chloroplast coupling factor 1.
The activation of the ATPase activity of coupling factor 1 (CF1) from chloroplasts by several detergents was studied. Further evidence that detergent micelles are important in the activation of Ca2+-ATPase was obtained. Maximal activation of CA2+-ATPase was achieved with short-chain alkyl-beta-D-glucopyranoside (alkylglucosides) detergents. Treatment of CF1 with hexylglucoside or heptylglucoside followed by hydroxylapatite chromatography caused nearly total removal of the epsilon subunit of the enzyme, whereas treatment with decylglucoside caused less ATPase activation and less loss of the epsilon subunit. The ATPase activity of detergent-activated CF1 was inhibited by purified epsilon subunit. Detergents that form small micelles appear to be most effective in removing the epsilon subunit and in activating the Ca2+-ATPase of CF1. When present during assay, the alkylglucosides also induce a Mg2+-ATPase activity in CF1. Octyl- and nonylglucoside are most effective in promoting this reaction. If, however, CF1 deficient in the epsilon subunit was used, even decylglucoside elicited rapid Mg2+-ATPase hydrolysis. It is concluded that removal of the epsilon subunit, although necessary for the expression of Mg2+-ATPase, is not sufficient. The detergents that cause maximal displacement of the epsilon subunit are less effective in inducing Mg2+-ATPase activity. The selective removal of subunits from CF1 by specific detergents points to potential problems with the use of these detergents in the solubilization of oligomeric membrane proteins.
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McCarty
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D000251:Adenosine Triphosphatases / Q000378:metabolism; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology*; D003902:Detergents / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005960:Glucosides / Q000494:pharmacology; D008823:Micelles; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013501:Surface-Active Agents / Q000494:pharmacology*; D013696:Temperature
D003902:Detergents; D005960:Glucosides; D008823:Micelles; D013501:Surface-Active Agents; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D006180:Proton-Translocating ATPases; D000252:Calcium-Transporting ATPases