Effect of crotapotin on the biological activity of Asp49 and Lys49 phospholipases A(2) from Bothrops snake venoms.
Myonecrosis, in addition to edema and other biological manifestations, are conspicuous effects of Bothrops snake venoms, some of them caused by phospholipases A(2) (PLA(2)s). Asp49-PLA(2)s are catalytically active, whereas Lys49-PLA(2)s, although highly toxic, have little or no enzymatic activity upon artificial substrates, due to a substitution of lysine for aspartic acid at position 49. Crotapotin (CA), the acidic counterpart of crotoxin PLA(2) (CB), is a PLA(2)-like protein from Crotalus durissus terrificus snake venom, and is considered a chaperone protein for CB, able to increase its lethality about ten fold, but to inhibit the formation of the rat paw edema induced by carrageenin and by snake venoms. In this study, we demonstrate that CA significantly inhibits the edema induced by BthTX-I (23% inhibition), BthTX-II (27%), PrTX-I (25%), PrTX-III (35%) and MjTX-II (10%) on the mouse paw. CK levels evoked by isolated Asp49 or Lys49-PLA(2)s were reduced by 40% to 54% in the presence of CA and, in all cases, the membrane damaging activity of the toxins was also reduced. Circular dichroism spectra of the PLA(2)s in the presence and absence of CA showed that there was not any detectable secondary structural modification due to association between CA and the myotoxins. However, Fourier Transformed Infrared (FT-IR) analysis indicated that ionic and hydrophobic contacts contributed to stabilize this interaction.
Cecchini
A L
AL
Depto de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, USP, Brazil.
Soares
A M
AM
Cecchini
R
R
de Oliveira
A H C
AH
Ward
R J
RJ
Giglio
J R
JR
Arantes
E C
EC
D000818:Animals; D017837:Bothrops; D002942:Circular Dichroism / Q000379:methods; D003402:Creatine Kinase / Q000378:metabolism; D003435:Crotalid Venoms / Q000201:enzymology*; D003439:Crotoxin / Q000032:analysis / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004487:Edema / Q000139:chemically induced / Q000473:pathology; D004791:Enzyme Inhibitors / Q000032:analysis / Q000633:toxicity*; D006614:Hindlimb; D008297:Male; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D010741:Phospholipases A / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000145:classification; D020033:Protein Isoforms; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods
D003435:Crotalid Venoms; D004791:Enzyme Inhibitors; D020033:Protein Isoforms; D003439:Crotoxin; D003402:Creatine Kinase; D010741:Phospholipases A