title: Purification and characterization of Cc-Lec, C-type lactose-binding lectin: A platelet aggregation and blood-clotting inhibitor from Cerastes cerastes venom.
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.
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Saoud
authors.initials: S
authors.lastname: Samah
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Chérifi
authors.initials: C
authors.lastname: Fatah
authors.affiliation: Poitiers University, Laboratory of Ecological and Biological Interactions, 15 Rue de l'Hôtel Dieu, Poitiers, France.
authors.forename: Berjeaud
authors.initials: B
authors.lastname: Jean-Marc
authors.affiliation: USTHB, Faculty of Chemistry, Laboratory of Theoretical Physico-Chemistry and Computer Chemistry, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Kellou-Taîri
authors.initials: KT
authors.lastname: Safia
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria. Electronic address: flaraba@hotmail.com.
authors.forename: Laraba-Djebari
authors.initials: LD
authors.lastname: Fatima
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology; D006384:Hemagglutination / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007785:Lactose / Q000378:metabolism*; D037181:Lectins, C-Type / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008670:Metals / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects*; D014757:Viper Venoms / Q000737:chemistry*
chemical_list: D037181:Lectins, C-Type; D008670:Metals; D014757:Viper Venoms; D007785:Lactose
keywords: Antiplatelet aggregation; C-type lectin; Cerastes cerastes; Coagulation; Homology modeling