Functions of novel serpin from Eudiplozoon nipponicum (Monogenea).
|Fakulta / Pracoviště MU|
|Popis||The properties of functional proteins of the members from Monogenea are still poorly investigated. We chose Eudiplozoon nipponicum as our experimental organism to address this issue. E. nipponicum (Diplozoidae, Polyopisthocotylea) is hematophagous ectoparasite which lives on the gills of common carp (Cyprinus carpio). The main aim of our current work is to understand the regulation of host/parasite peptidase activity related to numerous physiological processes. Among the key regulatory factors could be included the peptidase inhibitors, such as serpins - serine peptidase inhibitors. These functional proteins are generally known as important regulators of the coagulation cascade, complement, fibrinolysis, angiogenesis, inflammation etc. In the transcriptome of E. nipponicum we identified serpin gene (EnS), prepared it in recombinant form (rEnS) and investigated its properties. We have been able to achieve approx. 70% of protein sample purity. Using western blot, the presence of purified rEnS in bacterial extracts and EnS in excretory-secretory products (ESP) was confirmed. These results were validated by mass spectrometry (MS). Fluorometric inhibition assays showed the rEnS ability to partially inhibit four serine peptidases (SP) playing a role in host-parasite interaction – digestion (trypsin), regulation of blood coagulation (factor Xa, plasmin) or tempering the inflammation (kallikrein). Due to properties mentioned above and presence of the serpin in ESP, we hypothesize that EnS might be one of the key factor of host-parasite interaction.|