Antibodies against synthetic epitopes inhibit the enzymatic activity of mutalysin II, a metalloproteinase from bushmaster snake venom.
Mutalysin II (mut-II), a 22.5kDa zinc endopeptidase isolated from bushmaster (Lachesis muta muta) snake venom, is a direct acting fibrin(ogen)olytic proteinase. It induces monoclonal and polyclonal antibodies which efficiently neutralize the hemorrhagic effect of L. muta and several Bothrops whole venoms. To characterize epitopes of protective antibodies we have used the Spot method of multiple peptide synthesis to prepare 64 overlapping dodecapeptides frameshifted by three residues, covering the complete amino acid sequence of mut-II. The rabbit anti-mut-II antibodies binding pattern to peptides revealed several continuous antigenic regions: one in the N-terminal part, two in the central region and the other in the C-terminal of mut-II. By using homology modelling, a three-dimensional model of mut-II was built which showed that epitopes are surface exposed. Anti-peptide antibodies were raised against three peptides (one representative of each epitope region) covalently coupled as a mixture to keyhole limpet hemocyanin. Purified IgG from the resulting anti- peptide antibodies cross-reacted with mut-II and induced a dose-dependent inhibition of the mut-II catalyzed proteolysis of fibrinogen.
Ferreira
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RN
Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Machado de Avila
R A
RA
Sanchez
E F
EF
Maria
W S
WS
Molina
F
F
Granier
C
C
Chávez-Olórtegui
C
C
D000818:Animals; D000997:Antivenins / Q000494:pharmacology*; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D005260:Female; D007074:Immunoglobulin G / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000276:immunology; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D011817:Rabbits; D014757:Viper Venoms / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000276:immunology
D000997:Antivenins; D000939:Epitopes; D007074:Immunoglobulin G; D014757:Viper Venoms; D008666:Metalloendopeptidases; C068795:hemorrhagic factor LHFII