Masa-espektrometriaren bidezko giza-malkotako peptidoen analisia eta peptido antimikrobiarren identifikazioa

Abstract

Malkoak begia lubrifikatu eta babesten duten jariakinak dira. Bertan hainbat molekula desberdin daude, eta hauen artean peptido naturalak aurkitzen dira. Peptido hauek malkoaren babes-mekanismoen parte dira, izan ere aktibitate antimikrobiarra izan baitezakete. Malkoen peptido naturalen edukia sakonean karakterizatu gabe dirau gaur egun. Masa-espektrometria erabiliz, lan honetan malkotan inoiz identifikatu den peptido naturalen zerrenda handiena aurkezten da: 25 proteina desberdinetatik eratorritako 234 peptido. Identifikatutako bi peptidoren aktibitate antimikrobiarra frogatu da, erabilitako prozedura peptido antimikrobiarrak aurkitzeko eta karakterizatzeko baliogarria dela frogatuz. Abstract The tear film, an aqueous secretion protecting the eye, contains antimicrobial peptides involved in the protection against exogenous pathogens. In contrast to tryptic peptide studies, mass spectrometry analysis of endogenous peptides is difficult because of their heterogeneity. Therefore, we combined three mass spectrometry methods, leading to the generation of the biggest dataset of endogenous tear peptides to date. The antimicrobial activity prediction analysis performed in silico revealed different putative antimicrobial peptides among them. Two of the extracellular glycoprotein lacritin peptides were de novo synthesized, and their antimicrobial activity was confirmed in vitro. Our findings provide a useful approach for the discovery of peptides with antimicrobial activity.

The tear film, an aqueous secretion protecting the eye, contains antimicrobial peptides involved in the protection against exogenous pathogens. In contrast to tryptic peptide studies, mass spectrometry analysis of endogenous peptides is difficult because of their heterogeneity. Therefore, we combined three mass spectrometry methods, leading to the generation of the biggest dataset of endogenous tear peptides to date. The antimicrobial activity prediction analysis performed in silico revealed different putative antimicrobial peptides among them. Two of the extracellular glycoprotein lacritin peptides were de novo synthesized, and their antimicrobial activity was confirmed in vitro. Our findings provide a useful approach for the discovery of peptides with antimicrobial activity. Hitz gakoak: malkoak, peptidoak, peptidoma, peptido antimikrobiarrak, masa-espektrometria.