Purification and functional characterization of a new metalloproteinase (BleucMP) from Bothrops leucurus snake venom.
A fibrino(geno)lytic nonhemorrhagic metalloproteinase (BleucMP) was purified from Bothrops leucurus snake venom by two chromatographic steps procedure on DEAE-Sephadex A-25 followed by CM-Sepharose Fast Flow column. BleucMP represented 1.75% (w/w) of the crude venom and was homogeneous on SDS-PAGE. BleucMP analyzed by MALDI TOF/TOF, showed a molecular mass of 23,057.54Da and when alkylated and reduced, the mass is 23,830.40Da. Their peptides analyzed in MS (MALDI TOF\TOF) showed significant score when compared with those of other proteins by NCBI-BLAST2 alignment display. As regards their proteolytic activities, BleucMP efficiently acted on fibrinogen, fibrin, and was inhibited by EDTA and 1.10-phenanthroline. This enzyme was also able to decrease significantly the plasma fibrinogen level provoking blood incoagulability, however was devoid of hemorrhagic activity when tested in the mice skin and did not induce relevant biochemical, hematological and histopathological alterations in mice. The aspects addressed in this paper provide data on the effect of BleucMP in envenomation from B. leucurus snakes in order to better understand the effects caused by snake venom metalloproteinase.
Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia-MG, Brazil. mrochauesb@yahoo.com.br
Mário Sérgio R
MS
Gomes
Mayara R
MR
de Queiroz
Carla C N
CC
Mamede
Mirian M
MM
Mendes
Amélia
A
Hamaguchi
Maria I
MI
Homsi-Brandeburgo
Marcelo V
MV
Sousa
Elaine Nascimento
EN
Aquino
Mariana S
MS
Castro
Fábio
F
de Oliveira
Veridiana M
VM
Rodrigues
D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D002845:Chromatography / Q000379:methods; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology*; D005342:Fibrinolysis / Q000187:drug effects; D006470:Hemorrhage / Q000139:chemically induced / Q000473:pathology; D006801:Humans; D008297:Male; D008666:Metalloendopeptidases / Q000302:isolation & purification* / Q000633:toxicity; D051379:Mice; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000187:drug effects / Q000473:pathology; D010449:Peptide Mapping; D012867:Skin / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D018675:Toxicity Tests
D003435:Crotalid Venoms; D008666:Metalloendopeptidases