Antibodies to a fragment of the Bothrops moojenil-amino acid oxidase cross-react with snake venom components unrelated to the parent protein.
It is widely accepted that immunological cross-reactivity of snake venoms is mediated by antibodies that recognize venom components bearing either amino acid sequence homology or similar biological functions. However, here we demonstrate that polyspecific Bothrops antivenom is a source of cross-reactive antibodies that interact with venom proteins of distinctive primary structures and biological functions. The homoserine lactone derivative of the undecapeptide IQRWSLDKYAM (Ile1-Hse11), excised from the l-amino acid oxidase (LAAO) of the Bothrops moojeni venom, was the ligand of an affinity resin used to isolate specific anti-Ile1-Hse11 antibodies which were instrumental in revealing immunological cross-reactivity among unrelated venom proteins. We examined the extent of the cross-reactivity of these antibodies by probing electroblots of venoms from representative snakes of genera Bothrops, Lachesis, Crotalus and Micrurus, and by unambiguous structural characterization of the affinity-purified proteins of B. moojeni venom recovered from an agarose-anti-Ile1-Hse11 column. Our results indicate that all venoms tested had at least three reactive components toward anti-Ile1-Hse11 antibodies, among which we identified two serine proteases, one phospholipase A2 homologue, and LAAO. We hypothesize that the cross-reactivity of the anti-Ile1-Hse11 antibodies to unrelated venom proteins derives from their mechanism of antigen recognition, whereby complementarity is achieved through reciprocal conformational adaptation of the reacting molecules. Also, we believe these findings have implications both in the development of improved antivenoms and the preparation of immunochemical reagents for diagnostic and scientific investigation purposes in the field of snake venoms.
Stábeli
Rodrigo Guerino
RG
Departamento de Bioquímica e Imunologia, Faculdade de Medicina, USP, Ribeirão Preto, SP, Brazil.
Magalhães
Lívia Maria Pimenta
LM
Selistre-de-Araujo
Heloísa S
HS
Oliveira
Eduardo Brandt
EB
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000378:metabolism; D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D017837:Bothrops; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions / Q000276:immunology*; D003488:Cyanogen Bromide / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D050830:L-Amino Acid Oxidase; D020539:Sequence Analysis, Protein; D012910:Snake Venoms / Q000737:chemistry / Q000276:immunology* / Q000633:toxicity
D000911:Antibodies, Monoclonal; D012910:Snake Venoms; C088386:homoserine lactone; D000594:Amino Acid Oxidoreductases; D050830:L-Amino Acid Oxidase; D015107:4-Butyrolactone; D003488:Cyanogen Bromide