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Isolation and Functional Identification of an Antiplatelet RGD-Containing Disintegrin from Cerastes cerastes Venom

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Abstract

The current report focuses on purification, structural and functional characterization of Cerastategrin from Cerastes cerastes venom, a novel basic disintegrin (pI 8.36) with 128 amino acid residues and a molecular weight of 13 835.25 Da measured by MALDI-MSMS. The 3D structure of Cerastategrin is organized as α-helix (13%), β-strand (15%) and disordered structure (30%) and presents homologies with several snake venom disintegrins. Structural modeling shows that Cerastategrin presents an RGD motif that connects specifically to integrin receptors. Cerastategrin exhibits the inhibition of ADP induced platelets with an IC50 of 0.88 µg/mL and shows in vivo long stable anticoagulation effect 24 h post-injection of increasing doses ranging from 0.2 to 1 mg/kg, therefore, Cerastategrin maintained irreversibly the blood incoagulable. Moreover, Cerastategrin decreases the amount of bounded αIIbβ3 and reduced significantly the quantity of externalized P-Selectin. Cerastategrin acts as a molecule targeting specifically the receptor αIIbβ3; therefore, it behaves as a potent platelet activation inhibitor. As a new peptide with promising pharmacological properties, Cerastategrin could have a potential therapeutical effect in the vascular pathologies and may be a new effective treatment approach.

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Abbreviations

Cc1-PLA2 :

Cerastes cerastes Phospholipase A2

CCSV-MPase:

Cerastes cerastes Snake venom metalloproteinase

SDS PAGE:

Sodium dodecyl sulphate

H2SO4 :

Sulfuric acid

NaOH:

Sodium hydroxid

TFA:

Trifluorophosphate acide

ACN:

Acetonetrile

TCEP:

Tris(2-carboxyethyl) phosphine

IAM:

10 MM 2-iodoacetamide

EDTA:

Ethylenediaminetetraacetic

EGTA:

Aminopolycarboxylic acid

TRAP-6:

Thrombin receptor activator peptide-6

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Acknowledgements

The authors want to acknowledge Dr Hocine Rachid from INSERM UMRS 976 and University of Paris-Diderot, Saint Louis Hospital, Paris 75010, France for his help in purity control.

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Authors and Affiliations

  1. USTHB, Laboratory of Cellular and Molecular Biology, Faculty of Biological Sciences, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria

    Meriem Ameziani, Fatah Chérifi, Hamida Kiheli, Samah Saoud & Fatima Laraba-Djebari

  2. Faculty of Medicine, Blood Transfusion Center, Unverisity of Benyoucef Benkheda Algiers 1, CHU Bab El-Oued, Algiers, Algeria

    Ghania Hariti

  3. USTHB, Laboratory of Theoretical Physico-Chemistry and Computer Chemistry, Faculty of Chemistry, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria

    Safia Kellou-Taîri

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  1. Meriem Ameziani
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Ameziani, M., Chérifi, F., Kiheli, H. et al. Isolation and Functional Identification of an Antiplatelet RGD-Containing Disintegrin from Cerastes cerastes Venom. Protein J 39, 574–590 (2020). https://doi.org/10.1007/s10930-020-09915-y

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