Fractionation of membrane vesicles from coliphage M13-infected Escherichia coli.
Membrane vesicles were prepared by osmotic lysis of spheroplasts from M13-infected Escherichia coli. Reduced nicotinamide adenine dinucleotide (NADH) oxidase (reduced NAD: oxidoreductase, EC 1.6.99.3) and Mg2+-Ca2+-activated adenosine triphosphatase (ATP phosphohydrolase, EC 3.6.1.3), which are normally localized to the inner surface of the cytoplasmic membrane, were 50% acceesible to their polar substrates in these vesicles. The major coat protein of coliphage M13 is also bound to the cytoplasmic membrane (prior to phage assembly) but with its antigenic sites exposed to the exterior of the cell. Antibody to M13 coat protein was used to fractionate membrane vesicles. Neither agglutinated nor unagglutinated vesicles had altered NADH oxidase and adenosine triphosphatase specific activities. This is inconsistent with such vesicles being a mixture of correctly oriented and completely inverted membrane sacs and suggests that NADH oxidase, adenosine triphosphatase, M13 coat protein, or all three proteins rearrange during vesicle preparation.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D001666:Binding Sites, Antibody; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis / Q000201:enzymology / Q000648:ultrastructure; D003090:Coliphages / Q000032:analysis / Q000254:growth & development; D004267:DNA Viruses; D004926:Escherichia coli / Q000648:ultrastructure*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis
D014764:Viral Proteins; D009247:NADH, NADPH Oxidoreductases; D000251:Adenosine Triphosphatases