Role of the propeptide in folding and secretion of elastase of Pseudomonas aeruginosa.
Elastase of Pseudomonas aeruginosa is synthesized as a pre-proprotein. The propeptide has been shown to inhibit the enzymatic activity of elastase. In this study, we investigated a possible additional role of the propeptide in the folding and secretion of the enzyme. When elastase was expressed in Escherichia coli without its propeptide, no active elastase was produced. The enzyme was poorly released from the cytoplasmic membrane and, depending on the expression level, it was either degraded or it accumulated in an inactive form in the cell envelopes, probably as aggregates. Since proper folding is required for the release of translocated proteins from the cytoplasmic membrane and for the acquirement of a stable and active conformation, these results suggest that the propeptide is involved in the proper folding of the elastase and that it functions as an intramolecular chaperone. When mature elastase was expressed without its propeptide in P. aeruginosa, the enzyme was not secreted, and it was degraded. Therefore, proper folding of mature elastase appears to be required for secretion of the enzyme. Expression of the propeptide, as a separate polypeptide, in trans with mature elastase resulted in the formation of active elastase. This active enzyme was secreted in P. aeruginosa. Apparently, the propeptide can also function as an intermolecular chaperone.
Braun
P
P
Department of Molecular Cell Biology and Institute of Biomembranes, Utrecht University, Netherlands.
Tommassen
J
J
Filloux
A
A
D001426:Bacterial Proteins; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004789:Enzyme Activation; D004926:Escherichia coli / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D017510:Protein Folding; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D018832:Molecular Chaperones; D010455:Peptides; D011498:Protein Precursors; D008666:Metalloendopeptidases; C043859:pseudolysin, Pseudomonas aeruginosa