Snake venom metalloproteinases: structure/function relationships studies using monoclonal antibodies.
Snake Venom Metalloproteinases (SVMPs) are synthesized as zymogens and undergo proteolytic processing resulting in a variety of multifunctional proteins. Jararhagin is a P-III SVMP, isolated from the venom of Bothrops jararaca, comprising metalloproteinase, disintegrin-like and cysteine-rich domains. The catalytic domain is responsible for the hemorrhagic activity. The disintegrin-like/cysteine-rich domains block alpha2beta1 integrin binding to collagen and apparently enhance the hemorrhagic activity of SVMPs. The relevance of disintegrin-like domain is described in this paper using a series of mouse anti-jararhagin monoclonal antibodies (MAJar 1-7). MAJar 3 was the only antibody able to completely neutralize jararhagin hemorrhagic activity. Neutralization of catalytic activity was partial by incubation with MAJar 1. MAJars 1 and 3 efficiently neutralized jararhagin binding to collagen with IC50 of 330 and 8.4 nM, respectively. MAJars 1 and 3 recognized the C-terminal portion of the disintegrin domain, which is apparently in conformational proximity with the catalytic domain according to additivity tests. These data suggest that disintegrin-like domain epitopes are in close contact with catalytic site or functionally modulate the expression of hemorrhagic activity in SVMPs.
Tanjoni
Isabelle
I
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brasil, 1500, CEP 05503-900, São Paulo, SP, Brazil.
Butera
Diego
D
Bento
Luciana
L
Della-Casa
Maisa S
MS
Marques-Porto
Rafael
R
Takehara
Harumi A
HA
Gutiérrez
Jose M
JM
Fernandes
Irene
I
Moura-da-Silva
Ana M
AM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D017837:Bothrops; D003094:Collagen / Q000737:chemistry; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000276:immunology / Q000494:pharmacology*; D006470:Hemorrhage / Q000139:chemically induced; D008666:Metalloendopeptidases / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D045726:Metalloproteases / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D013329:Structure-Activity Relationship; D000097146:Bothrops jararaca Venom
D000911:Antibodies, Monoclonal; D003094:Collagen; D003435:Crotalid Venoms; D008666:Metalloendopeptidases; D045726:Metalloproteases; D000097146:Bothrops jararaca Venom