Naslov
VaSP1, catalytically active serine proteinase from Vipera ammodytes ammodytes venom with unconventional active site triad

Autori
Kurtović, Tihana ; Brgles, Marija ; Leonardi, Adrijana ; Lang Balija, Maja ; Sajevic, Tamara ; Križaj, Igor ; Allmaier, Günter ; Marchetti-Deschmann, Martina ; Halassy, Beata

Izvornik
Toxicon (0041-0101) 77 (2014); 93-104

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
snake venom serine proteinase; non-canonical catalytic triad; Vipera ammodytes; mass spectrometry

Sažetak
VaSP1, a serine proteinase from Vipera ammodytes ammodytes venom, is a glycosylated monomer of 31.5 kDa, as determined by MALDI mass spectrometry, showing multiple isoelectric points between pH 6.5 and pH 8.5. Partial amino acid sequencing of VaSP1 by Edman degradation and MS/MS analysis identified sequences which allowed its classification among the so-called snake venom serine proteinase homologues, members of the peptidase S1 family, however being devoid of the canonical catalytic triad. Only few representatives of this group have been identified so far with just two of them characterised in detail at the protein level. Despite substitution of His57 with Arg, VaSP1 possesses proteolytic activity which can be inhibited by Pefabloc, benzamidine, Zn2+ ions, DTT and trypsin inhibitor II, a Kunitz/BPTI group member. It hydrolyses Nα-benzoyl-Phe-Val-Arg-p-NA, exhibiting Michaelis-Menten behaviour with Km = 48.2 µM and Vm = 0.019 nM s-1. The pH for optimal activity on tested substrate is around 9.0. VaSP1 also cleaves insulin B-chain, digesting it at positions His10-Leu11, Ala14-Leu15 and Tyr16-Leu17. Furthermore, the novel serine proteinase is active towards wide array of proteins involved in hemostasis where its degradation of fibrinogen, fibrin, prothrombin, factor X and plasminogen in vivo probably results in depletion of coagulation factors in blood circulation. The possibility that VaSP1 possesses anticoagulant properties has been further indicated by its ability to prolong prothrombin time and activated partial thromboplastin time.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija

POVEZANOST RADA