title: Maturation of Pseudomonas aeruginosa elastase. Formation of the disulfide bonds.
abstract: 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 in dsb 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.
authors.affiliation: Department of Molecular Cell Biology, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Braun
authors.forename: C
authors.initials: C
authors.lastname: Ockhuijsen
authors.forename: E
authors.initials: E
authors.lastname: Eppens
authors.forename: M
authors.initials: M
authors.lastname: Koster
authors.forename: W
authors.initials: W
authors.lastname: Bitter
authors.forename: J
authors.initials: J
authors.lastname: Tommassen
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D010196:Pancreatic Elastase / Q000737:chemistry / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013440:Sulfides
chemical_list: D001426:Bacterial Proteins; D013440:Sulfides; D010196:Pancreatic Elastase