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 essential step for colonization and infection. In most known cases, attachment is mediated by a specific protein-carbohydrate interaction. We have used a carbohydrate-containing crosslinking probe to select bacterial surface adhesins for trypsin digestion, MALDI-TOF mass spectrometry and identification against genome sequence. The present paper describes this functional proteomics approach for identification of the recently cloned low-abundant Lewis(b)-binding adhesin of Helicobacter pylori. Protein identification was obtained through the enrichment of approximately 300 fmol of adhesin from solubilized cells.
Larsson
T
T
Institute of Medical Biochemistry, Göteborg University, Box 440, SE-405 30, Göteborg, Sweden.
Bergström
J
J
Nilsson
C
C
Karlsson
K A
KA
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006028:Glycosphingolipids / Q000737:chemistry*; D016480:Helicobacter pylori / Q000737:chemistry*; D006801:Humans; D012709:Serum Albumin / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D019809:Streptavidin / Q000737:chemistry
D018829:Adhesins, Bacterial; D006028:Glycosphingolipids; C024050:Le(b)-active glycolipid; D012709:Serum Albumin; D019809:Streptavidin