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Biotinylation and Purification of Surface-exposed Helicobacter pylori Proteins.

Bio-protocol (2015-10-09)
Bradley J Voss, Timothy L Cover
ABSTRACT

Interactions between pathogenic bacteria and host cells are often mediated by proteins found on the surfaces of the bacteria. The Gram-negative bacterium Helicobacter pylori is predicted to produce at least 50 surface-exposed outer membrane proteins, but there has been relatively little progress in experimentally analyzing the cell-surface proteome of this organism. Herein, we describe in detail a protocol that allows biotinylation and purification of surface-exposed H. pylori proteins. A comparative analysis of surface-exposed proteins identified by this biotinylation-based approach and by several other independent methods is described in a recent publication (Voss et al., 2014).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
Sigma-Aldrich
Biotin, ≥99% (HPLC), lyophilized powder
Sigma-Aldrich
Trichloroacetic acid solution, 6.1 N
Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt dihydrate, ACS reagent, 99.0-101.0%