Reconstitution of D-Lactate Dehydrogenase-Deficient Membrane Vesicles using Fluorine-labeled Components: An Approach to Investigating Protein-Lipid Interactions in Biological Membranes

As part of an investigation of structure-function relationships in biological membranes by biochemical and biophysical methods, the authors have been studying the membrane-bound enzyme D-lactate dehydrogenase (D-LDH) of Escherichia coli , and the effects on its activity of varying the lipid composit...

Full description

Saved in:
Bibliographic Details
Published in:Biophysical journal 1982-01, Vol.37 (1), p.101-103
Main Authors: Pratt, E A, Jones, J A, Cottam, P F, Dowd, S R, Ho, C
Format: Article
Language:English
Subjects:
D-lactate dehydrogenase
Escherichia coli
membrane vesicles
Poster Summaries
reconstitution
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:As part of an investigation of structure-function relationships in biological membranes by biochemical and biophysical methods, the authors have been studying the membrane-bound enzyme D-lactate dehydrogenase (D-LDH) of Escherichia coli , and the effects on its activity of varying the lipid composition of the membrane. Because fluorine-19 can be a useful probe for nuclear magnetic resonance (NMR) studies, the authors have incorporated 8,8-difluoromyristic acid into the lipids of unsaturated fatty acid auxotrophs, and have prepared membrane vesicles containing super(19)F-labeled lipids. In addition, the authors can incorporate 4-, 5-, and 6-fluorotryptophans into the proteins of E. coli W3110trp A33 and extract super(19)F-labeled D-LDH. These super(19)F-labeled components have been tested for their ability to reconstitute D-LDH-dependent activity in D-LDH-deficient membrane vesicles.
ISSN:0006-3495
1542-0086
DOI:10.1016/S0006-3495(82)84621-3