BaltDC: purification, characterization and infrared spectroscopy of an antiplatelet DC protein isolated from Bothrops alternatus snake venom.

Autor: Matias MS; Postgraduate Program in Genetics and Biochemistry, Institute of Genetics and Biochemistry, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil., de Sousa BB; Postgraduate Program in Genetics and Biochemistry, Institute of Genetics and Biochemistry, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil.; National Institute of Science and Technology in Nanobiopharmaceutics (N-Biofar), Belo Horizonte, MG Brazil., da Cunha Pereira DF; Postgraduate Program in Genetics and Biochemistry, Institute of Genetics and Biochemistry, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil., Dias EHV; Postgraduate Program in Genetics and Biochemistry, Institute of Genetics and Biochemistry, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil., Mamede CCN; Institute of Agricultural Sciences, Federal University of Uberlândia (UFU), Monte Carmelo, MG Brazil.; National Institute of Science and Technology in Nanobiopharmaceutics (N-Biofar), Belo Horizonte, MG Brazil., de Queiroz MR; National Institute of Science and Technology in Nanobiopharmaceutics (N-Biofar), Belo Horizonte, MG Brazil., Silva ACA; Institute of Physics, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil., Dantas NO; Institute of Physics, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil., Soares AM; Center for the Study of Biomolecules Applied to Health (CEBio), Oswaldo Cruz Foundation (Fiocruz - Rondônia) and Health Group, Federal University of Rondônia (UNIR), Porto Velho, RO Brazil.; University Center São Lucas (UniSL), Porto Velho, RO Brazil., de Oliveira Costa J; Postgraduate Program in Genetics and Biochemistry, Institute of Genetics and Biochemistry, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil.; Federal Institute of Education, Science and Technology of Triângulo Mineiro (IFTM), Campus Ituiutaba, Ituiutaba, MG Brazil., de Oliveira F; Institute of Biomedical Sciences, Federal University of Uberlândia (UFU), Uberlândia, MG Brazil.; National Institute of Science and Technology in Nanobiopharmaceutics (N-Biofar), Belo Horizonte, MG Brazil.
Jazyk: angličtina
Zdroj: The journal of venomous animals and toxins including tropical diseases [J Venom Anim Toxins Incl Trop Dis] 2017 Jul 28; Vol. 23, pp. 36. Date of Electronic Publication: 2017 Jul 28 (Print Publication: 2017).
DOI: 10.1186/s40409-017-0126-7
Abstrakt: Background: 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 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.
Methods: The 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.
Results: BaltDC 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 PO 3 2- groups, present in BaltDC, form hydrogen bonds with the PO 2 - groups present in the non-lipid portion of the membrane platelets.
Conclusions: BaltDC may be of medical interest since it was able to inhibit platelet aggregation.
Databáze: MEDLINE
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