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Infection and Immunity, July 2002, p. 3396-3403, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3396-3403.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Proteome Analysis of Secreted Proteins of the Gastric Pathogen Helicobacter pylori

Dirk Bumann,1 Sevil Aksu,2 Meike Wendland,1 Katharina Janek,3 Uschi Zimny-Arndt,2 Nicolas Sabarth,1 Thomas F. Meyer,1* and Peter R. Jungblut2

Department of Molecular Biology,1 Central Core Facility for Protein Analysis, Max-Planck-Institute for Infection Biology,2 Charité, Institute for Biochemistry, Berlin, Germany3

Received 5 December 2001/ Returned for modification 14 January 2002/ Accepted 21 March 2002

Secreted proteins (the secretome) of the human pathogen Helicobacter pylori may mediate important pathogen-host interactions, but such proteins are technically difficult to analyze. Here, we report on a comprehensive secretome analysis that uses protein-free culture conditions to minimize autolysis, an efficient recovery method for extracellular proteins, and two-dimensional gel electrophoresis followed by peptide mass fingerprinting for protein resolution and identification. Twenty-six of the 33 separated secreted proteins were identified. Among them were six putative oxidoreductases that may be involved in the modification of protein-disulfide bonds, three flagellar proteins, three defined fragments of the vacuolating toxin VacA, the serine protease HtrA, and eight proteins of unknown function. A cleavage site for the amino-terminal passenger domain of VacA between amino acids 991 and 992 was determined by collision-induced dissociation mass spectrometry. Several of the secreted proteins are interesting targets for antimicrobial chemotherapy and vaccine development.

* Corresponding author. Mailing address: Max-Planck-Institut für Infektionsbiologie, Abteilung Molekulare Biologie, Schumannstraße 21/22, D-10117 Berlin, Germany. Phone: 49 30 28460 400. Fax: 49 30 28460 401. E-mail: meyer{at}mpiib-berlin.mpg.de.

Editor: E. I. Tuomanen

Infection and Immunity, July 2002, p. 3396-3403, Vol. 70, No. 7
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.7.3396-3403.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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