ABSTRACT
A new secretory phospholipase A2 (sPLA2) isoform from Bothrops jararacussu venom (BjVIII) has been characterized by causing platelet aggregation, an absent activity in BthTx-I, Prtx-I and PrTx-II sPLA2s. According to our results, BjVIII also enhances insulin release by the pancreatic beta cells. The complete amino acid sequence of the new isoform was determined by Edman degradation and de novo peptide sequencing. These analyses showed a G35K amino acid modification for BjVIII in comparison with BthTx-I, PrTx-I and Prtx-II, a structural difference that has been related to the conflicting biological activities among BjVIII and other Lys49 sPLA2s. The whole set of evidences collected in this work indicates that, besides the C-terminal region and B-wing of PLA2, the calcium binding loop in BjVIII should be considered as an important region, involved in the pharmacological effects of Lys49-sPLA2 isoforms from the Bothrops genus.
Subject(s)
Biocatalysis , Bothrops , Crotalid Venoms/enzymology , Insulin-Secreting Cells/metabolism , Insulin/metabolism , Lysine , Phospholipases A2, Secretory/pharmacology , Amino Acid Sequence , Animals , Insulin Secretion , Insulin-Secreting Cells/drug effects , Isoenzymes/chemistry , Isoenzymes/isolation & purification , Isoenzymes/metabolism , Isoenzymes/pharmacology , Molecular Sequence Data , Phospholipases A2, Secretory/chemistry , Phospholipases A2, Secretory/isolation & purification , Phospholipases A2, Secretory/metabolism , Rats , Structure-Activity RelationshipABSTRACT
BjVIII is a new myotoxic Lys49-PLA2 isolated from Bothrops jararacussu venom that exhibits atypical effects on human platelet aggregation. To better understand the mode of action of BjVIII, crystallographic studies were initiated. Two crystal forms were obtained, both containing two molecules in the asymmetric unit (ASU). Synchrotron radiation diffraction data were collected to 2.0 A resolution and 1.9 A resolution for crystals belonging to the space group P2(1)2(1)2(1) (a = 48.4 A, b = 65.3 A, c = 84.3 A) and space group P3(1)21 (a = b = 55.7 A, c = 127.9 A), respectively. Refinement is currently in progress and the refined structures are expected to shed light on the unusual platelet aggregation activity observed for BjVIII.
