Solubilization and partial purification of alanine carrier from membranes of a thermophilic bacterium and its reconstitution into functional vesicles. |
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Authors: | H Hirata N Sone M Yoshida Y Kagawa |
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Institution: | Department of Biochemistry, Jichi Medical School, Minamikawachi-machi, Tochigi-ken, Japan 329-04 |
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Abstract: | An alanine transport carrier was solubilized from membranes of the thermophilic bacterium PS3 with cholate-deoxycholate mixture. It was then partially purified by diethyl aminoethyl cellulose column chromatography and gel filtration. For assay of alanine carrier activity it was reconstituted into vesicles with P-lipids and the transport energy was supplied as a membrane potential introduced by K+-diffusion via valinomycin. The partially purified carrier had no ATPase or NADH dehydrogenase activity. Active transport of alanine driven by the membrane potential was completely abolished by an uncoupler. |
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