Maturation of Pseudomonas aeruginosa Elastase

Elastase of Pseudomonas aeruginosa is synthesized as a preproenzyme. After propeptide-mediated folding in the periplasm, the proenzyme is autoproteolytically processed, prior to translocation of both the mature enzyme and the propeptide across the outer membrane. The formation of the two disulfide b...

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Bibliographic Details
Published inThe Journal of biological chemistry Vol. 276; no. 28; pp. 26030 - 26035
Main Authors Braun, Peter, Ockhuijsen, Corrine, Eppens, Elaine, Koster, Margot, Bitter, Wilbert, Tommassen, Jan
Format Journal Article
LanguageEnglish
Published Elsevier Inc 13.07.2001
American Society for Biochemistry and Molecular Biology
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Summary:Elastase of Pseudomonas aeruginosa is synthesized as a preproenzyme. After propeptide-mediated folding in the periplasm, the proenzyme is autoproteolytically processed, prior to translocation of both the mature enzyme and the propeptide across the outer membrane. The formation of the two disulfide bonds present in the mature enzyme was examined by studying the expression of the wild-type enzyme and of alanine for cysteine mutant derivatives in the authentic host and indsb mutants of Escherichia coli. It appeared that the two disulfide bonds are formed successively. First, DsbA catalyzes the formation of the disulfide bond between Cys-270 and Cys-297 within the proenzyme. This step is essential for the subsequent autoproteolytic processing to occur. The second disulfide bond between Cys-30 and Cys-57 is formed more slowly and appears to be formed after processing of the proenzyme, and its formation is catalyzed by DsbA as well. This second disulfide bond appeared to be required for the full proteolytic activity of the enzyme and contributes to its stability.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M007122200