Site-directed mutagenesis of Glu-141 and His-223 in Pseudomonas aeruginosa elastase: catalytic activity, processing, and protective activity of the elastase against Pseudomonas infection.

Infection and Immunity
S KawamotoK Okuda

Abstract

Both Pseudomonas aeruginosa elastase and Bacillus thermoproteolyticus thermolysin are zinc metalloproteases. On the basis of the high homology of the P. aeruginosa elastase with the Bacillus thermolysin, we hypothesized that Glu-141 and His-223 are the key residues for catalytic activity of the Pseudomonas elastase. To test this possibility, we replaced Glu-141 with Asp, Gln, and Gly and His-223 with Gly, Glu, and Leu by site-directed mutagenesis. These substitutions dramatically diminished the proteolytic activities of the mutant elastases when they were expressed in Escherichia coli cells. Although these mutant elastase precursors (proelastases) were produced, no appreciable processing was observed with these mutants. The possibility that autocatalysis is involved in both the processing and activation of elastase is discussed. Furthermore, by immunizing mice with vaccines made from these mutant elastase, we were able to obtain good protection against an intraperitoneal P. aeruginosa challenge.

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Feb 1, 1992·Molecular Microbiology·M J Brumlik, D G Storey
Jan 1, 1989·Antibiotics and Chemotherapy·K MoriharaK Okuda
Dec 1, 1989·Molecular Microbiology·C Wandersman
Apr 1, 1985·Canadian Journal of Microbiology·T I Nicas, B H Iglewski
Jun 1, 1987·Journal of Bacteriology·P A SchadB H Iglewski
Jan 1, 1986·Journal of Clinical Microbiology·K Morihara, J Y Homma
May 16, 1988·Biochemical and Biophysical Research Communications·S YamamotoK Morihara
Nov 1, 1988·Journal of Bacteriology·E Kessler, M Safrin
Jan 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·T A Kunkel
Jul 12, 1972·Nature: New Biology·B W MatthewsD Dupourque
Jul 12, 1972·Nature: New Biology·B W MatthewsH Neurath

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Citations

Feb 11, 2005·Journal of Biomolecular Structure & Dynamics·Olayiwola A AdekoyaIngebrigt Sylte
Aug 24, 2006·Molecular Microbiology·Pamela J BonnerLawrence J Shimkets
Dec 1, 2006·FEMS Microbiology Letters·Hui YangYun Li
Oct 25, 1996·The Journal of Biological Chemistry·M J O'Donohue, A Beaumont
Jun 22, 2005·Journal of Bacteriology·C KooiP A Sokol
Jul 10, 2007·The Journal of Immunology : Official Journal of the American Association of Immunologists·Sukumar SahaKenji Okuda
Apr 16, 1998·The Journal of Biological Chemistry·C Marie-ClaireA Beaumont
Oct 30, 2007·Journal of Bacteriology·Alan Pavinski BitarHélène Marquis
Apr 25, 2006·Médecine et maladies infectieuses·R Le BerreB Guery
Dec 1, 1993·Microbiological Reviews·C C Häse, R A Finkelstein
Nov 7, 1996·Chemical Reviews·William N. Lipscomb, Norbert Sträter
Mar 28, 2009·Biochimie·Tania Yu GromovaSergey V Kostrov

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