Structural properties of the proteoliposomes catalyzing electron transport from formate to fumarate.
The electron-transport chain catalyzing fumarate reduction by formate has recently been reconstituted from the formate dehydrogenase complex and the fumarate reductase complex from Vibrio succinogenes, in a liposomal preparation containing vitamin K-1 (Unden, G. and Kröger, A. (1982) Biochim. Biophys. Acta 682, 258-263). We have now investigated the structural properties of this preparation. The preparation was found to consist of a homogeneous population of unilamellar proteoliposomes with an average diameter of about 100 nm and an internal volume of 2-4 ml/g phospholipid. The buoyant density (1.07 g/ml) was consistent with the protein/phospholipid ratio (0.2 g/g) of the preparation. Leakage of glucose from the internal spaces of the proteoliposomes was negligibly slow. Proteoliposomes prepared with either of the enzyme complexes showed peripheral projections mainly on the outer surface, when examined by electron microscopy after negative staining. The size, orientation and surface density of the projections were consistent with those of the enzymes. Most of the substrate and dye-reactive sites (70-90%) of the enzymes in the proteoliposomes were accessible to external non-permeant substrates. The proteoliposomes catalyzing electron transport were formed by freeze-thawing a mixture of liposomes and protein-phospholipid complexes which did not perform electron transport from formate to fumarate. Nearly the entire amount of the enzymes supplied (0.2 g protein/g phospholipid) was incorporated into the liposomes by this procedure. The transformation of liposomes into proteoliposomes was accompanied by exchange of the internal solutes with the external medium.
D000445:Aldehyde Oxidoreductases / Q000378:metabolism*; D004579:Electron Transport; D005560:Formate Dehydrogenases / Q000378:metabolism*; D005561:Formates / Q000378:metabolism*; D005650:Fumarates / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D008854:Microscopy, Electron; D009097:Multienzyme Complexes / Q000378:metabolism*; D011510:Proteolipids / Q000378:metabolism*; D013385:Succinate Dehydrogenase / Q000378:metabolism*; D014733:Vibrio / Q000201:enzymology*
D005561:Formates; D005650:Fumarates; D008081:Liposomes; D009097:Multienzyme Complexes; D011510:Proteolipids; C020485:proteoliposomes; C030544:formic acid; C032005:fumaric acid; D005560:Formate Dehydrogenases; D000445:Aldehyde Oxidoreductases; C033727:fumarate reductase complex; D013385:Succinate Dehydrogenase