[en] <title>Abstract</title>
<p>Proteomic characterization of snake venoms is essential for understanding the molecular basis of their evolution and for identifying bioactive compounds of therapeutic interest. The <italic>Montivipera</italic> species endemic to the Near and Middle East region remain poorly studied despite their interesting biological activities. Previous analyses of <italic>Montivipera</italic> venoms have revealed only partial proteomic profiles, with notable discrepancies between studies. To address this gap, we conducted a proteomic analysis of five <italic>Montivipera</italic> species, including <italic>M. bornmuelleri</italic>, <italic>M. bulgardaghica</italic>, <italic>M. albizona</italic>, <italic>M. raddei</italic> and <italic>M. xanthina</italic>. We also analyzed the venom of <italic>Macrovipera lebetinus</italic> to provide a broader comparative framework. These venoms were analyzed using an integrated approach combining SDS-PAGE, RP-HPLC and shotgun proteomics, using both trypsin and multi-enzymatic limited digestions to maximize protein identification and coverage. SDS-PAGE and RP-HPLC analyses revealed the remarkable complexity and diversity of <italic>Montivipera</italic> venoms, which were further confirmed by shotgun proteomics, identifying between 129 and 179 proteins and peptides per species. The major protein families detected included snake venom metalloproteinases, phospholipases A<sub>2</sub>, venom serine proteases, C-type lectins, venom vascular-endothelial growth factors, and disintegrins. Notably, the relative abundance of these protein families varied across species, suggesting interspecific differences in envenomation profiles. Comparative analysis revealed a high degree of similarity among <italic>Montivipera</italic> species, with 39 shared proteins across all five venoms. Our findings confirmed the major toxin families previously reported in <italic>Montivipera</italic> venoms and revealed the presence of several low-abundance protein families that were not previously identified. Thus, this study highlights both the conserved and unique features of <italic>Montivipera</italic> venom proteomes, offering a valuable foundation for future functional and evolutionary investigations.</p>
