Skp, a molecular chaperone of gram-negative bacteria, is required for the formation of soluble periplasmic intermediates of outer membrane proteins.
Using a cross-linking approach, we have analyzed the function of Skp, a presumed molecular chaperone of the periplasmic space of Escherichia coli, during the biogenesis of an outer membrane protein (OmpA). Following its transmembrane translocation, OmpA interacts with Skp in close vicinity to the plasma membrane. In vitro, Skp was also found to bind strongly and specifically to pOmpA nascent chains after their release from the ribosome suggesting the ability of Skp to recognize early folding intermediates of outer membrane proteins. Pulse labeling of OmpA in spheroplasts prepared from an skp null mutant revealed a specific requirement of Skp for the release of newly translocated outer membrane proteins from the plasma membrane. Deltaskp mutant cells are viable and show only slight changes in the physiology of their outer membranes. In contrast, double mutants deficient both in Skp and the periplasmic protease DegP (HtrA) do not grow at 37 degrees C in rich medium. We show that in the absence of an active DegP, a lack of Skp leads to the accumulation of protein aggregates in the periplasm. Collectively, our data demonstrate that Skp is a molecular chaperone involved in generating and maintaining the solubility of early folding intermediates of outer membrane proteins in the periplasmic space of Gram-negative bacteria.
Schäfer
U
U
Institut für Biochemie und Molekularbiologie, Universität Freiburg, Hermann-Herderstrasse 7, D-79104 Freiburg, Germany.
Beck
K
K
Müller
M
M
D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D017353:Gene Deletion; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009675:Novobiocin / Q000494:pharmacology; D019897:Periplasm / Q000378:metabolism; D033903:Periplasmic Proteins; D011485:Protein Binding; D017510:Protein Folding; D012293:Rifampin / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000235:genetics; D013696:Temperature; D014640:Vancomycin / Q000494:pharmacology
D001425:Bacterial Outer Membrane Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D033903:Periplasmic Proteins; C072721:Skp protein, E coli; C078167:OMPA outer membrane proteins; D009675:Novobiocin; D014640:Vancomycin; C061682:DegP protease; D012697:Serine Endopeptidases; D012293:Rifampin