Thus, trimarin may play a key role in the pathophysiological conditions that occur during T. Remarkably, the enzyme clotted the factor X deficient human plasma, suggesting that trimarin has factor Xa-like activity. Trimarin coagulated the citrated human plasma in the absence of CaCl 2, but it was lacking thrombin like activity as it did not clot the purified fibrinogen. Trimarin shortened the activated partial thromboplastin time (aPTT) and prothrombin time (PT), suggesting its involvement in common pathway of blood coagulation. Trimarin showed procoagulant activity with reduced re-calcification time of citrated human plasma. Myotoxicity due to muscle tissue necrosis was substantiated with increased serum CK activity. The hemorrhage and myotoxicity correlated with degradation of extracellular protein components type-IV collagen and fibronectin. Trimarin exhibited pharmacological activities including hemorrhage, myotoxicity, procoagulant and factor Xa-like activities. 28 Intravital microscopy was conducted on an upright microscope (Model 512815/20 Leitz, Wetzlar, Germany) with a saline immersion objective (SW 40/0.75 numerical aperture). Bars represent the mean ± SD of five individual experiments. 74 LEITZ WETZLAR LAMP 307-127. Anesthesia and surgical preparation of the cremaster muscle for intravital microscopy experiments was performed as described previously. CK analysis in serum: (B) Blood from the respective animals were collected by cardiac puncture after anesthesia, serum was assayed for CK level in units/l. The metal ion associated with trimarin was found to be Zn 2+. The sections were stained with hematoxyline-eosin and pictures were taken under Leitz wetzlar Germany type-307-148.002 microscope. Trimarin showed proteolytic activity towards casein and fibrinogen, which was irreversibly inhibited by EDTA and 1,10-phenanthroline. Trimarin is a single-chain basic protein, with a molecular mass of 29.6 kDa.
In the present study, we describe the purification and characterization of a metalloprotease ‘trimarin’ from Trimeresurus malabaricus snake venom.
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