Cloning, expression, and hemostatic activities of a disintegrin, r-mojastin 1, from the mohave rattlesnake (Crotalus scutulatus scutulatus).
Interactions with exposed subendothelial extracellular proteins and cellular integrins (endothelial cells, platelets and lymphocytes) can cause alterations in the hemostatic system associated with atherothrombotic processes. Many molecules found in snake venoms induce pathophysiological changes in humans, cause edema, hemorrhage, and necrosis. Disintegrins are low molecular weight, non-enzymatic proteins found in snake venom that mediate changes by binding to integrins of platelets or other cells and prevent binding of the natural ligands such as fibrinogen, fibronectin or vitronectin. Disintegrins are of great biomedical importance due to their binding affinities resulting in the inhibition of platelet aggregation, adhesion of cancer cells, and induction of signal transduction pathways. RT-PCR was used to obtain a 216 bp disintegrin cDNA from a C. s. scutulatus snake venom gland. The cloned recombinant disintegrin called r-mojastin 1 codes for 71 amino acids, including 12 cysteines, and an RGD binding motif. r-Mojastin 1 inhibited platelet adhesion to fibronectin with an IC50 of 58.3 nM and ADP-induced platelet aggregation in whole blood with an IC50 of 46 nM. r-Mojastin 1 was also tested for its ability to inhibit platelet ATP release using PRP resulting with an IC50 of 95.6 nM. MALDI-TOF mass spectrum analysis showed that r-mojastin has a mass of 7.95676 kDa.
Sánchez
Elda E
EE
Natural Toxins Research Center, College of Arts and Sciences, 975 W. Avenue B. MSC 158, Texas A&M University-Kingsville, Kingsville, TX 78363, USA.
Lucena
Sara E
SE
Reyes
Steven
S
Soto
Julio G
JG
Cantu
Esteban
E
Lopez-Johnston
Juan Carlos
JC
Guerrero
Belsy
B
Salazar
Ana Maria
AM
Rodríguez-Acosta
Alexis
A
Galán
Jacob A
JA
Tao
W Andy
WA
Pérez
John C
JC
D000255:Adenosine Triphosphate / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000201:enzymology*; D017839:Crotalus; D003545:Cysteine; D019483:Disintegrins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000378:metabolism; D006487:Hemostasis / Q000187:drug effects*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D003435:Crotalid Venoms; D019483:Disintegrins; D005353:Fibronectins; D009842:Oligopeptides; D011994:Recombinant Proteins; C554078:mojastin protein, Crotalus scutulatus; C047981:arginyl-glycyl-aspartic acid; D000255:Adenosine Triphosphate; D003545:Cysteine