Snake venomics of the Armenian mountain vipers Macrovipera lebetina obtusa and Vipera raddei Libia Sanz a , Naira Ayvazyan b , Juan J. Calvete a, ⁎ a Instituto de Biomedicina de Valencia, C.S.I.C., Valencia, Spain b Department of Biophysics, Yerevan State University, Armenia ARTICLE INFO ABSTRACT Article history: Received 7 May 2008 Accepted 26 May 2008 Venoms from the Armenian mountain vipers Macrovipera lebetina obtusa and Vipera raddei were analyzed by RP-HPLC, N-terminal sequencing, MALDI-TOF mass fingerprinting and CID-MS/ MS. The venom proteins of M.l. obtusa and V. raddei belong to 9 and 11 families, respectively. The two mountain viper venoms share bradykinin-potentiating/C-natriuretic peptides, and proteins from the dimeric distegrin, DC-fragment, CRISP, PLA 2 , serine proteinase, C-type lectin- like, L-amino acid oxidase, and Zn 2+ -dependent metalloproteinase families, albeit each species exhibits distinct relative abundances. M.l. obtusa and V. raddei venoms contain unique components, e.g. the short disintegrin obtustatin in M.l. obtusa, and Kunitz-type serine proteinase inhibitor and VEGF-like molecules in V. raddei. The toxin formulation of M.l. obtusa and V. raddei venoms may be related to their adaptation to rocky mountain ecosystems. On the other hand, the possibility that the VEGF-like proteins from V. raddei underlie the reported potential therapeutic value of V. raddei venom for regenerating damaged peripheral nerves deserves further investigations. Using a similarity coefficient, we estimate that the similarity of venom proteins between M. l. obtusa and M. l. transmediterranea is less than 4%. Although this result would support the classification of M.l. obtusa and M.l. transmediterranea as different species, additional detailed genomic analyses are also required. © 2008 Elsevier B.V. All rights reserved. Keywords: Snake venomics Macrovipera lebetina obtusa Vipera raddei Snake venom protein families Proteomics Mass spectrometry 1. Introduction Venom toxins, produced by a pair of specialized glands in the upper jaw [1,2], likely evolved from proteins with a normal physiological function that were recruited into the venom proteome before the diversification of the advanced snakes, at the base of the Colubroidea radiation [3–6], and represent the critical innovation that allowed advanced snakes to transition from a mechanical (constriction) to a chemical (venom) means of subduing and digesting prey larger than themselves. Venom proteins have multiple functions including immobilizing, paralyzing, killing and digesting prey. Venoms produced by snakes of the family Viperidae (vipers and pit vipers) contain proteins that interfere with the coagulation cascade, the normal haemostatic system and tissue repair, and human envenoma- tions are often characterized by clotting disorders, hypofibrino- genemia and local tissue necrosis [7]. In spite of the fact that viperid venoms may contain well over 100 protein components [8], venom proteins belong to only a few major protein families, including enzymes (serine proteinases, Zn 2+ -metalloprotei- nases, L-amino acid oxidase, group II PLA 2 ) and proteins without enzymatic activity (disintegrins, C-type lectins, natriuretic JOURNAL OF PROTEOMICS 71 (2008) 198 – 209 ⁎ Corresponding author. Instituto de Biomedicina de Valencia, C.S.I.C., Jaime Roig 11, 46010 Valencia, Spain. Tel.: +34 96 339 1778; fax: +34 96 369 0800. E-mail address: jcalvete@ibv.csic.es (J.J. Calvete). 1874-3919/$ – see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.jprot.2008.05.003 available at www.sciencedirect.com www.elsevier.com/locate/jprot