Mitra Jyotirmoy, Bhattacharyya Debasish
Division of Structural Biology and Bioinformatics, CSIR-Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Jadavpur, Kolkata, West Bengal 700032, India.
Division of Structural Biology and Bioinformatics, CSIR-Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Jadavpur, Kolkata, West Bengal 700032, India.
Toxicon. 2014 Sep;88:1-10. doi: 10.1016/j.toxicon.2014.06.004. Epub 2014 Jun 14.
Phosphodiesterases (PDEs) belong to a super-family of enzymes that have multiple roles in the metabolism of extracellular nucleotides and regulation of nucleotide-based intercellular signalling. A PDE from Russell's viper (Daboia russelli russelli) venom (DR-PDE) was purified by gel filtration, ion exchange and affinity chromatographies. Homogeneity of the preparation was verified by SDS-PAGE, SE-HPLC and mass spectrometry. It was free from 5'-nucleotidase, alkaline phosphatase and protease activities. Identity of the enzyme was ensured from partial sequence homology with other PDEs. DR-PDE was inactivated by polyvalent anti-venom serum and metal chelators. The enzyme was partially inhibited by the root extracts of four medicinal plants but remained unaffected by inhibitors of intracellular PDEs. DR-PDE hydrolyses ADP and thus, strongly inhibits ADP-induced platelet aggregation in human platelet rich plasma. This study leads to better understanding of a component of Russell's viper venom that affects homoeostatic system of the victim.
磷酸二酯酶(PDEs)属于一类酶的超家族,它们在细胞外核苷酸代谢和基于核苷酸的细胞间信号调节中具有多种作用。通过凝胶过滤、离子交换和亲和色谱法纯化了罗素蝰蛇(Daboia russelli russelli)毒液中的一种磷酸二酯酶(DR-PDE)。通过SDS-PAGE、尺寸排阻高效液相色谱(SE-HPLC)和质谱法验证了制剂的纯度。它没有5'-核苷酸酶、碱性磷酸酶和蛋白酶活性。通过与其他磷酸二酯酶的部分序列同源性确定了该酶的身份。DR-PDE被多价抗蛇毒血清和金属螯合剂灭活。该酶被四种药用植物的根提取物部分抑制,但不受细胞内磷酸二酯酶抑制剂的影响。DR-PDE水解ADP,因此强烈抑制富含人血小板血浆中ADP诱导的血小板聚集。这项研究有助于更好地理解罗素蝰蛇毒液中影响受害者体内稳态系统的一种成分。