Crystallization and preliminary X-ray diffraction studies of piratoxin III, a D-49 phospholipase A₂ from the venom of Bothrops pirajai

Abstract

Piratoxin III (PrTX-III) is a phospholipase A 2 (PLA 2 , E.C. 3.1.1.4, phosphatide sn-2 acylhydrolase) isolated from Bothrops pirajai. Crystals of PrTX-III were obtained using the vapour-diffusion technique and X-ray diffraction data have been collected to 2.7 A Ê resolution. The enzyme was crystallized in the space group C2 with unit-cell parameters a = 60.88, b = 100.75, c = 48.19 A Ê , = 123.89 . A molecular-replacement solution of the structure has been found using bothropstoxin I from the venom of B. jararacussu as a search model.

Renal and cardiovascular effects of Bothrops marajoensis venom and phospholipase A2

“…sPLA 2 activity was measured following the methods described by Wen-Hwa et al (1999) and modified by Toyama et al (2003) for 96-well plates using 4-nitro-3-octanoyloxy-benzoic acid as a substrate (4N3OBA, BIOMOL, USA). Enzyme activity, expressed as the initial velocity of the reaction (Vo), was calculated based on the increase in absorbance after 20 min.…”

Harpalycin 2 inhibits the enzymatic and platelet aggregation activities of PrTX-III, a D49 phospholipase A2 from Bothrops pirajai venom

“…sPLA2 activity was measured following the protocols described by Lee et al [13] and modified by Toyama et al [12] for 96-well plate, using 4-nitro-3-octanoyloxy-benzoic acid (4N3OBA, manufactured by BIOMOL, USA) as the substrate. Enzyme activity was calculated based on the increase in absorbance after 20 min.…”

“…Exogenous administration of sPLA 2 , such as snake venom sPLA 2 , induces and/or exacerbates inflammatory response in animals [11,12]. Structural analyses revealed that snake venom sPLA 2 s have a similar molecular profile to those of human secretory PLA 2 s as well as a conserved catalytic site [13], thus making them useful tools for the search of new anti-phospholipase A 2 drugs.…”

Role of phospholipase A2 and tyrosine kinase in Clostridium difficile toxin A‐induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion

“…PLA 2 activity was measured following the protocol described by Wen-Hwa et al (1999) and modi¼ed by Toyama et al (2003) for a 96well plate. The standard assay mixture contained 200 μl of buffer (10 mM Tris-HCl, 10 mM CaCl2, 100 mM and NaCl, pH 7.8), 20 μl of substrate [4-nitro-3-octanoyloxybenzoic acid (NOBA)], 20 μl of water and 20 μl of PLA 2 , giving a ¼nal volume of 260 μl.…”