Specific interaction between OutD, an Erwinia chrysanthemi outer membrane protein of the general secretory pathway, and secreted proteins.
OutD is an outer membrane component of the main terminal branch of the general secretory pathway (GSP) in Erwinia chrysanthemi. We analyzed the interactions of OutD with other components of the GSP (Out proteins) and with secreted proteins (PelB, EGZ and PemA). OutD is stabilized by its interaction with another GSP component, OutS. The 62 C-terminal amino acids of OutD are necessary for this interaction. In vivo formation of OutD multimers, up to tetramers, was proved after the dissociation in mild conditions of the OutD aggregates formed in the outer membrane. Thus, OutD could form a channel-like structure in the outer membrane. We showed that OutD is stabilized in vivo when co-expressed with Out-secreted proteins. This stabilization results from the formation of complexes that were detected in experiments of co-immunoprecipitation and co-sedimentation in sucrose density gradients. The presence of the N-terminal part of OutD is required for this interaction. The interaction between OutD and the secreted protein PelB was confirmed in vitro, suggesting that no other component of the GSP is required for this recognition. No interaction was observed between the E. carotovora PelC and the E. chrysanthemi OutD. Thus, the interaction between GspD and the secreted proteins present in the periplasm could be the key to the specificity of the secretion machinery and a trigger for that process.
Shevchik
V E
VE
Laboratoire de Génétique Moléculaire des Microorganismes et des Interactions Cellulaires, CNRS UMR 5577, INSA, Villeurbanne, France. shevchik@insa.insa-lyon.fr
Robert-Baudouy
J
J
Condemine
G
G
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D001692:Biological Transport; D002462:Cell Membrane / Q000648:ultrastructure; D002480:Cellulase / Q000378:metabolism; D016974:Dickeya chrysanthemi / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D008958:Models, Molecular; D016296:Mutagenesis; D016973:Pectobacterium carotovorum / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017384:Sequence Deletion; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D007527:Isoenzymes; C106887:OutD protein, Erwinia chrysanthemi; C106888:OutS protein, Erwinia chrysanthemi; C080692:endoglucanase Z; D002480:Cellulase; D011133:Polysaccharide-Lyases; C074124:pectin lyase B; C023911:pectate lyase