Česká společnost pro hmotnostní spektrometrii

(2. konference České společnosti pro hmotnostní spektrometrii - WeP-011)
Nepenthesins – carnivorous plant proteases as a tool for hydrogen/deuterium exchange mass spectrometry

Alan Kádek ^1,2 *, Petra Darebná ², Petr Halada ¹, Petr Novák ^1,2, Petr Man ^1,2

1. Institute of Microbiology, Academy of Sciences of the Czech Republic
2. Faculty of Science, Charles University in Prague, Czech Republic

Abstrakt

Pitcher plants (genus Nepenthes - láčkovky) are carnivorous plants inhabiting tropical areas of southeastern Asia – mainly Borneo and Sumatra. Their prey-trapping mechanism is based on a leave-derived conical organ called “pitcher” used for trapping and digestion of insects. Digestive fluids present in the pitcher have been shown to contain several interesting enzymes, among which acidic aspartic proteases called nepenthesins are especially appealing for their potential use in hydrogen/deuterium exchange (HDX) mass spectrometry.

In this work we enriched nepenthesins from the pitcher fluid of multiple Nepenthes species and used them for protein digestion in HDX workflow. We compared digestion efficiency and cleavage specificity of nepenthesins with pepsin, a gold standard for protein digestion in HDX.

* Korespondující autor: kadek@biomed.cas.cz

Poděkování:

The authors thank Dr. Vlastik Rybka from Prague Botanical Garden in Troja for the access to nepenthes collection and valuable advice. This work has been financially supported by the Grant Agency of the Czech Republic (GACR P206/12/0503) and by the Institutional Research Project of the Institute of Microbiology (RVO61388971).