A biochemical study of the reconstitution of d-lactate dehydrogenase-deficient membrane vesicles using fluorine-labeled components |
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Authors: | E.A. Pratt Judith A. Jones Patricia F. Cottam Susan R. Dowd Chien Ho |
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Affiliation: | Department of Biological Sciences, Carnegie-Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213 U.S.A. |
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Abstract: | Fluorine-19 labeled compounds have been incorporated into lipids and proteins of Escherichia coli. 19F-Labeled membrane vesicles, prepared by growing a fatty acid auxotroph of a d-lactate dehydrogenase-deficient strain on 8,8-difluoromyristic acid, can be reconstituted for oxidase and transport activities by binding exogenous d-lactate dehydrogenase. 19F-Labeled d-lactate dehydrogenases prepared by addition of fluorotryptophans to a tryptophan-requiring strain are able to reconstitute d-lactate dehydrogenase-deficient membrane vesicles. Thus, lipid and protein can be labeled independently and used to investigate protein-lipid interactions in membranes. |
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Keywords: | Reconstitution Membrane vesicle Protein-lipid interaction (E. coli) MTT 3-(4,5-dimethylthiazolyl-2)2,5-diphenyl-tetrazolium bromide PMS phenazine methosulfate |
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