Peptides from Pisum sativum L. enzymatic protein digest with anti-adhesive activity against Helicobacter pylori: structure-activity and inhibitory activity against BabA, SabA, HpaA and a fibronectin-binding adhesin.
METHODS
Identification of anti-adhesive peptides against Helicobacter pylori obtained by enzymatic hydrolysis of seed proteins from Pisum sativum L. (Fabaceae).


RESULTS
Bioassay-guided fractionation of protein tryptic digest by ultrafiltration, size exclusion chromatography (SEC) and reversed phase chromatography (RPC) were used. Identification of bioactive peptides was achieved by MALDI-TOF-MS. Adhesion of H. pylori was monitored by two different assays, using a quantitative in vitro assay on human AGS cells with evaluation of bacterial binding by flow cytometry, beside a semi-quantitative in situ adhesion assay using FITC-labelled H. pylori on human stomach tissue sections. From two highly active fractions (F3, F3.3) two anti-adhesive peptides (S3, S5) were identified. Neither F3 nor S3 or S5 had any cytotoxic effect against H. pylori. By hemagglutination assay and semiquantitative dot blot overlay assay with immobilized ligands it was shown that F3 interacts specifically with H. pylori adhesins BabA, SabA, HpaA and a fibronectin-binding adhesin, while S3 and S5 inhibit only BabA. It was demonstrated that BabA, usually interacting with carbohydrate motifs such as fucosylated blood group antigens, interacts with the peptide moieties.


CONCLUSIONS
Bioactive peptides from pea protein could be applied as functional ingredients for protecting infants and children against infections such as H. pylori.
Institute for Pharmaceutical Biology and Phytochemistry, University of Münster, Münster, Germany.
Michael
M
Niehues
Marco
M
Euler
Gilda
G
Georgi
Marko
M
Mank
Bernd
B
Stahl
Andreas
A
Hensel
D018829:Adhesins, Bacterial / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001422:Bacterial Adhesion / Q000187:drug effects*; D002460:Cell Line; D005353:Fibronectins / Q000378:metabolism; D016480:Helicobacter pylori / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D018532:Pisum sativum / Q000737:chemistry*; D010455:Peptides / Q000494:pharmacology*; D010940:Plant Proteins / Q000737:chemistry; D011485:Protein Binding; D012639:Seeds / Q000737:chemistry; D013329:Structure-Activity Relationship
D018829:Adhesins, Bacterial; D000900:Anti-Bacterial Agents; C400429:BabA protein, Helicobacter pylori; D005353:Fibronectins; D010455:Peptides; D010940:Plant Proteins; C500808:SabA protein, Helicobacter pylori; C095977:adhesin A, Helicobacter