title: Identification of surface proteins of Helicobacter pylori by selective biotinylation, affinity purification, and two-dimensional gel electrophoresis.
abstract: Helicobacter pylori is a widespread human pathogen that can cause gastric ulcers and cancer. To identify surface proteins that may play a role in pathogen-host interactions and represent potential targets for the control of this infection, we selectively biotinylated intact H. pylori with the hydrophilic reagent sulfosuccinimidyl-6-(biotinamido)-hexanoate and purified the labeled proteins by membrane isolation, solubilization, and affinity chromatography. After separation of 82 biotinylated proteins on two-dimensional gels, 18 were identified with comparison to proteome data and peptide mass fingerprinting. Among the identified proteins, 9 have previously been shown to be surface-exposed, 7 are associated with virulence, and 11 are highly immunogenic in infected patients. In conclusion, this generally applicable combined proteome approach facilitates the rapid identification of promising targets for the control of H. pylori and might be applicable to numerous other human pathogens although larger biotinylation reagents might be required in some cases to prevent permeation of porin channels in the outer membrane.
authors.affiliation: Max-Planck-Institut für Infektionsbiologie, Abteilung Molekulare Biologie, Schumannstrabetae 21/22, D-10117 Berlin, Germany.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Sabarth
authors.forename: Stefanie
authors.initials: S
authors.lastname: Lamer
authors.forename: Ursula
authors.initials: U
authors.lastname: Zimny-Arndt
authors.forename: Peter R
authors.initials: PR
authors.lastname: Jungblut
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Meyer
authors.forename: Dirk
authors.initials: D
authors.lastname: Bumann
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D019921:Biotinylation; D015180:Electrophoresis, Gel, Two-Dimensional; D004798:Enzymes / Q000737:chemistry / Q000302:isolation & purification; D016480:Helicobacter pylori / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D004798:Enzymes; D008565:Membrane Proteins