Use of an affinity proteomics approach for the identification of low-abundant bacterial adhesins as applied on the Lewis(b)-binding adhesin of Helicobacter pylori

Microbial attachment to host cell surfaces is considered to be the first es sential step for colonization and infection. In most known cases, attachmen t is mediated by a specific protein-carbohydrate interaction. We have used a carbohydrate-containing crosslinking probe to select bacterial surface ad hesins for trypsin digestion, MALDI-TOF mass spectrometry and identificatio n against genome sequence. The present paper describes this functional prot eomics approach for identification of the recently cloned low-abundant Lewi s(b)-binding adhesin of Helicobacter pylori. Protein identification was obt ained through the enrichment of approximately 300 fmol of adhesin from solu bilized cells. (C) 2000 Federation of European Biochemical Societies.