J Med Microbiol Email Content Delivery
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Dassanayake, R. P.
Right arrow Articles by Duhamel, G. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dassanayake, R. P.
Right arrow Articles by Duhamel, G. E.
Agricola
Right arrow Articles by Dassanayake, R. P.
Right arrow Articles by Duhamel, G. E.
J Med Microbiol 53 (2004), 319-323; DOI: 10.1099/jmm.0.45549-0
© 2004 Society for General Microbiology
ISSN 0022-2615

Biochemical properties of membrane-associated proteases of Brachyspira pilosicoli isolated from humans with intestinal disorders

Rohana P. Dassanayake1, Nancy E. Caceres2, Gautam Sarath2,3 and Gerald E. Duhamel1,2

Department of Veterinary and Biomedical Sciences1, Center for Biotechnology2 and Department of Biochemistry3, University of Nebraska-Lincoln, Lincoln, NE 68583-0905, USA

Correspondence Gerald E. Duhamel gduhamel1{at}unl.edu, Gautam Sarath gsarath1{at}unl.edu

Received September 19, 2003
Accepted December 3, 2003

A membrane-associated, subtilisin-like, serine protease activity was found in both pathogenic and non-pathogenic strains of Brachyspira species in a previous study, but the biochemical properties of the enzyme were not investigated. The purpose of the present study was to characterize further the biochemical properties, including substrate specificity, of the membrane-associated protease of Brachyspira pilosicoli isolated from humans with intestinal disorders. Protease activity of detergent-enriched membrane protein extracts of B. pilosicoli was assessed using fluorescent dye-labelled synthetic peptides as substrates and determination of electrophoretic mobility of cleavage products in agarose gels. Each activity was further confirmed with class-specific protease inhibitors and thermal denaturation. The presence of a hydrophilic membrane-associated thermolabile serine endopeptidase with specificity for Leu was confirmed. Two additional hydrophilic membrane-associated thermostable proteolytic activities were identified, one with a putative Ala specificity, and one a carboxypeptidase. Taken together, these data suggest that, in addition to a previously described membrane-associated subtilisin-like serine protease, the membrane of B. pilosicoli contains proteins with at least two other proteolytic activities.

This paper was presented at the Second International Conference on Colonic Spirochaetal Infections in Animals and Humans, Edinburgh, UK, 2–4 April 2003.

Abbreviation: ODG, n-octyl ß-D-glucopyranoside.




This article has been cited by other articles:


Home page
 Antimicrob. Agents Chemother.Home page
R. P. Dassanayake, G. Sarath, and G. E. Duhamel
Penicillin-Binding Proteins in the Pathogenic Intestinal Spirochete Brachyspira pilosicoli
Antimicrob. Agents Chemother., April 1, 2005; 49(4): 1561 - 1563.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
INT J SYST EVOL MICROBIOL J MED MICROBIOL MICROBIOLOGY J GEN VIROL ALL SGM JOURNALS
Copyright © 2004 Society for General Microbiology.