Abstract
BackgroundSnake venoms are a complex mixture of proteins, organic and inorganic compounds. Some of these proteins, enzymatic or non-enzymatic ones, are able to interact with platelet receptors, causing hemostatic disorders. The possible therapeutic potential of toxins with antiplatelet properties may arouse interest in the pharmacological areas. The present study aimed to purify and characterize an antiplatelet DC protein from Bothrops alternatus snake venom.MethodsThe protein, called BaltDC (DC protein from B. alternatus snake venom), was purified by a combination of ion-exchange chromatography on DEAE-Sephacel column and gel filtration on Sephadex G-75. The molecular mass was estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). The amino acid sequence of the N-terminal region was carried out by Edman degradation method. Platelet aggregation assays were performed in human platelet-rich plasma (PRP). Infrared (IR) spectroscopy was used in order to elucidate the interactions between BaltDC and platelet membrane.ResultsBaltDC ran as a single protein band on SDS-PAGE and showed apparent molecular mass of 32 kDa under reducing or non-reducing conditions. The N-terminal region of the purified protein revealed the amino acid sequence IISPPVCGNELLEVGEECDCGTPENCQNECCDA, which showed identity with other snake venom metalloproteinases (SVMPs). BaltDC was devoid of proteolytic, hemorrhagic, defibrinating or coagulant activities, but it showed a specific inhibitory effect on platelet aggregation induced by ristocetin and epinephrine in PRP. IR analysis spectra strongly suggests that PO32− groups, present in BaltDC, form hydrogen bonds with the PO2− groups present in the non-lipid portion of the membrane platelets.ConclusionsBaltDC may be of medical interest since it was able to inhibit platelet aggregation.
Highlights
Snake venoms are a complex mixture of proteins, organic and inorganic compounds
We describe the purification, characterization and IR spectra of an antiplatelet DC protein, called BaltDC, isolated from B. alternatus snake venom
We described the purification, electrophoretic profile, N-terminal sequence and IR spectra of an antiplatelet DC protein from this snake venom
Summary
Snake venoms are a complex mixture of proteins, organic and inorganic compounds. Some of these proteins, enzymatic or non-enzymatic ones, are able to interact with platelet receptors, causing hemostatic disorders. The non-protein fraction of the venom is composed by low-mass organic molecules, such as carbohydrates, lipids, free amino acids and inorganic compounds including calcium, phosphorus, magnesium, potassium, sodium and zinc [1, 2]. D and C domains are linked by disulfide bonds, so they are released as a unique molecule (DC protein) This molecule is able to interact with integrins present on the cell surface and may trigger numerous cellular processes such as platelet aggregation; angiogenesis; metastasis; tumor growth; adhesion, migration and proliferation of cells [8,9,10,11,12]
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More From: Journal of Venomous Animals and Toxins including Tropical Diseases
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