Purification and characterization of tenerplasminin-1, a serine peptidase inhibitor with antiplasmin activity from the coral snake ( Micrurus tener tener ) venom

Abstract

A plasmin inhibitor, named tenerplasminin-1 (TP1), was isolated from Micrurus tener tener (Mtt) venom. It showed a molecular mass of 6542 Da, similarly to Kunitz-type serine peptidase inhibitors. The amidolytic activity of plasmin (0.5 nM) on synthetic substrate S-2251 was inhibited by 91% following the incubation with TP1 (1 nM). Aprotinin (2 nM) used as the positive control of inhibition, reduced the plasmin amidolytic activity by 71%. Plasmin fibrinolytic activity (0.05 nM) was inhibited by 67% following incubation with TP1 (0.1 nM). The degradation of fibrinogen chains induced by plasmin, trypsin or elastase was inhibited by TP1 at a 1:2, 1:4 and 1:20 enzyme:inhibitor ratio, respectively. On the other hand, the proteolytic activity of crude Mtt venom on fibrinogen chains, previously attributed to metallopeptidases, was not abolished by TP1. The tPA-clot lysis assay showed that TP1 (0.2 nM) acts like aprotinin (0.4 nM) inducing a delay in lysis time and lysis rate which may be associated with the inhibition of plasmin generated from the endogenous plasminogen activation. TP1 is the first serine protease plasmin-like inhibitor isolated from Mtt snake venom which has been characterized in relation to its mechanism of action, formation of a plasmin:TP1 complex and therapeutic potential as anti-fibrinolytic agent, a biological characteristic of great interest in the field of biomedical research. They could be used to regulate the fibrinolytic system in pathologies such as metastatic cancer, parasitic infections, hemophilia and other hemorrhagic syndromes, in which an intense fibrinolytic activity is observed.

Snake Venom Peptides: Tools of Biodiscovery

“…It is a slow and tight binding inhibitor of plasmin. The peptide (TP1) isolated from Micrurus tener tener venom is also an inhibitor of plasmin [ 39 , 121 , 122 ]. TP1 has a molecular mass similar to Kunitz-type peptides, but its sequence is not reported till now.…”

“…The peptide and its complex also inhibited the fibrinogen-clotting activity of thrombin. Another Kunitz-type inhibitor (namely PIVL) isolated from Macrovipera lebetina venom, displayed potent anti-tumor activity through interference with integrin receptor function [ 26 , 122 , 123 , 124 , 125 ].…”

Contribution of endothelial cell and macrophage activation in the alterations induced by the venom of Micrurus tener tener in C57BL/6 mice

“…The simultaneous release of pro-inflammatory mediators, such as TNF-α, may promote PAI-1 expression, thus inhibiting this activity (Levi and van der Poll, 2017;Petäjä, 2011). In addition, the low pro-fibrinolytic activity may be associated with tenerplasminin-1, a serine protease inhibitor isolated from Mtt venom that decreases the activity of plasmin and delays clot lysis induced by tPA (Vivas et al, 2016). Since binding to cell receptors confers protection to plasminogen activators from PAI-1 inhibition (Li et al, 2007), our next approach was evaluating pro-fibrinolytic activity on the cell surface.…”

“…Micrurus envenomation is characterized by neurotoxic symptoms and may lead to death from muscle paralysis and respiratory arrest. Some reports have shown that these venoms may also contain hemorrhagic, hemostatic, hemolytic and edematogenic activities (Barros et al, 1994;Casais-e-Silva and Teixeira, 2017;Salazar et al, 2011Salazar et al, , 2018Tambourgi et al, 1994;Vivas et al, 2016). In the United States, only two species are responsible for all coral snake toxicity, Micrurus fulvius fulvius (Eastern coral snake) and Micrurus tener tener (Texas coral snake), and most of the information available rely on the Eastern coral snakebites.…”

Toxicosis of Snake, Scorpion, Honeybee, Spider, and Wasp Venoms: Part 1

“…Cerestes cerestes antivenom and Macrovipera mauritanica antivenom cross react with Bitis arietans antivenom due to presence of antigens common to them [31]. Tenerplasminin-1 (TP 1 ), a plasmin inhibitor isolated from Micrurus tener tener venom was similar to Kunitztype serine peptidase inhibitors [32]. Rauwolfia serpentina inhibits Daboia russelii venom [33].…”