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Purification and characterization of Cc-Lec, C-type lactose-binding lectin: A platelet aggregation and blood-clotting inhibitor from Cerastes cerastes venom

Author:
Samah, Saoud, Fatah, Chérifi, Jean-Marc, Berjeaud, Safia, Kellou-Taîri, Fatima, Laraba-Djebari
Source:
International journal of biological macromolecules 2017 v.102 pp. 336-350
ISSN:
0141-8130
Subject:
adenosine diphosphate, affinity chromatography, amino acids, arachidonic acid, calcium, coagulation, crosslinking, disulfide bonds, erythrocytes, fibrinogen, hemagglutination, humans, lectins, mass spectrometry, mice, platelet aggregation, polyacrylamide gel electrophoresis, polypeptides, receptors, therapeutics, thrombosis, venoms
Abstract:
In this study, we reported for the first time the biochemical and structural characterization of Cc-Lec, a C-type lectin purified from Cerastes cerastes venom by affinity chromatography. This lectin was homogeneous by SDS-PAGE, and was shown to be a 34 271.59Da polypeptide by Electrospray mass spectrometry MS-ES-TOF. Its identified sequence of 160 amino acids corresponding to one subunit, revealed a high identity with other related proteins. Cc-Lec modeled 3D structure appeared as homodimer cross-linked by one disulfide bridge. Cc-Lec exhibited a calcium dependent hemagglutinating activity against human group O erythrocytes. Cc-Lec inhibited platelet aggregation induced by ADP, arachidonic acid or fibrinogen suggesting its interaction with their specific receptors namely P2Y1 and/or P2Y12, GPIIb/IIIa and TPα respectively. Cc-Lec was not lethal for mice until 10mg/kg administered by i.p. route. The lectin displayed a lasting anticoagulation on mice plasma even two days post-injection. This anticoagulation seems to be related to its interaction with coagulation factors Xa and IXa. Therefore, Cc-Lec prevented FXa amidolytic activity with Km=4.3310−4μg/mL and ki=14.4μg/mL. It seems to interact with these targets through CRD domain which could make it a good target as a pharmacological promising molecule in thrombosis diagnosis and therapy.
Agid:
5666967