title: Biochemical properties of a new PI SVMP from Bothrops pauloensis: inhibition of cell adhesion and angiogenesis.
abstract: In the present work, we demonstrate some biochemical and functional properties of a new PI snake venom metalloproteinase (SVMP) isolated from Bothrops pauloensis snake venom (BpMP-II), in addition we evaluated its capacity to inhibit endothelial cell adhesion and in vitro angiogenesis. BpMP-II was purified after a combination of three chromatography steps and showed molecular mass of 23,000 Da determined by MALDI-TOF, an isoelectric point of 6.1 and the sequence of some fragments obtained by MS/MS (MALDI TOF\TOF) presented high structural similarity with other PI-SVMPs. BpMP-II showed proteolytic activity against azocasein, was able to degrade bovine fibrinogen and was inhibited by EDTA, 1.10 phenantroline and β-mercaptoethanol. BpMP-II did not induce local hemorrhage in the dorsal region of mice even at high doses and did not affect plasma creatine kinase (CK) levels when administered intramuscularly into the gastrocnemius muscle of mice. Moreover, this metalloproteinase decreased tEnd cells viability at concentrations higher than 20 μg/mL. With sub-toxic doses this metalloproteinase affected tEnd cell adhesion and was also able to inhibit in vitro angiogenesis. BpMP-II showed very important functional properties suggesting considerable therapeutic potential for this class of protein.
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil.
authors.forename: David Collares
authors.initials: DC
authors.lastname: Achê
authors.affiliation: Department of Chemical and Physical, State University of Southwest Bahia (UESB), 45506-210 Jequie, BA, Brazil.
authors.forename: Mário Sérgio R
authors.initials: MS
authors.lastname: Gomes
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil.
authors.forename: Dayane Lorena Naves
authors.initials: DL
authors.lastname: de Souza
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil.
authors.forename: Makswell Almeida
authors.initials: MA
authors.lastname: Silva
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil.
authors.forename: Maria Inês Homsi
authors.initials: MI
authors.lastname: Brandeburgo
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil.
authors.forename: Kelly Aparecida Geraldo
authors.initials: KA
authors.lastname: Yoneyama
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil.
authors.forename: Renata Santos
authors.initials: RS
authors.lastname: Rodrigues
authors.affiliation: Ezequiel Dias Foundation, FUNED, 30510-010 Belo Horizonte, MG, Brazil.
authors.forename: Márcia Helena
authors.initials: MH
authors.lastname: Borges
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil. Electronic address: lsdaiana@yahoo.com.br.
authors.forename: Daiana Silva
authors.initials: DS
authors.lastname: Lopes
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, 38400-902 Uberlandia, MG, Brazil. Electronic address: veridiana@ingeb.ufu.br.
authors.forename: Veridiana de Melo
authors.initials: Vde M
authors.lastname: Rodrigues
mesh_terms: D000818:Animals; D017837:Bothrops; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D042783:Endothelial Cells / Q000187:drug effects; D005342:Fibrinolysis / Q000187:drug effects; D007797:Laminin / Q000737:chemistry; D045726:Metalloproteases / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011509:Proteoglycans / Q000737:chemistry; D012910:Snake Venoms / Q000737:chemistry / Q000201:enzymology*
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D012910:Snake Venoms; C059485:matrigel; D003094:Collagen; D045726:Metalloproteases
keywords: Angiogenesis inhibition; Bothrops pauloensis; Endothelial cells; Matrigel; Metalloproteinase; Snake venom