A biochemical study of the reconstitution of D-lactate dehydrogenase-deficient membrane vesicles using fluorine-labeled components.
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.
D002462:Cell Membrane / Q000378:metabolism*; D004926:Escherichia coli; D005227:Fatty Acids / Q000032:analysis; D005456:Fluorescent Dyes / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009227:Myristic Acids / Q000378:metabolism; D014364:Tryptophan / Q000031:analogs & derivatives / Q000378:metabolism
D005227:Fatty Acids; D005456:Fluorescent Dyes; D008563:Membrane Lipids; D008565:Membrane Proteins; D009227:Myristic Acids; C026927:5-fluorotryptophan; C001244:6-fluorotryptophan; C037847:8,8-difluoromyristic acid; D014364:Tryptophan; D007770:L-Lactate Dehydrogenase