M13 procoat inserts into liposomes in the absence of other membrane proteins.
Procoat, the precursor form of the major coat protein of coliphage M13, assembles into the Escherichia coli inner membrane and is cleaved to mature coat protein by leader peptidase. This assembly process has previously been reconstituted using lipids and purified leader peptidase in a cell-free protein synthesis reaction (Watts, C., Silver, P., and Wickner, W. (1981) Cell 25, 347-353; Ohno-Iwashita, Y., and Wickner, W. (1983) J. Biol. Chem. 258, 1895-1900). We now report that procoat can also cross a liposomal membrane composed of only purified phospholipids; leader peptidase is not needed to catalyze insertion. When procoat is synthesized in vitro in the presence of liposomes with encapsulated chymotrypsin, the procoat inserts spontaneously through the membrane and is degraded. The protease was shown by several criteria to be in the lumen of the liposomes. These results demonstrate that the precursor form of an E. coli integral membrane protein can cross a membrane without the aid of leader peptidase or any other membrane proteins.
D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D002918:Chymotrypsin / Q000378:metabolism; D003090:Coliphages; D019286:Endopeptidase K; D010450:Endopeptidases / Q000378:metabolism / Q000502:physiology; D004926:Escherichia coli / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D008565:Membrane Proteins / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011498:Protein Precursors / Q000378:metabolism*; D012697:Serine Endopeptidases
D036022:Capsid Proteins; D008081:Liposomes; D008565:Membrane Proteins; D009842:Oligopeptides; D010743:Phospholipids; D011480:Protease Inhibitors; D011498:Protein Precursors; C025668:coat protein, Bacteriophage M13; C002101:chymostatin; D010450:Endopeptidases; D012697:Serine Endopeptidases; D002918:Chymotrypsin; D019286:Endopeptidase K; C026515:type I signal peptidase