title: Purification and variability in thrombin-like activity of Bothrops atrox venom from different geographic regions.
abstract: Bothrops atrox snake venoms from two different Amazon regions, i.e., Manaus, AM (3 degrees 0.6'40"S; 60 degrees 0.1'6.0"W) and Tucurui, PA (3 degrees 0.42'30"S; 49 degrees 0.41'45"W), were analyzed with respect to the thrombin-like activity component by elution profile on gel-filtration and reverse phase HPLC chromatography, electrophoretic mobility on SDS-PAGE, and enzymatic activity on fibrinogen. Despite some individual discrepancies among venom specimens, the thrombin-like activity present in the Manaus pool was eluted earlier compared with the Tucurui pool but its enzymatic specific activity on thrombin was lower (s.a. = 6.0) than that observed in the Tucurui pool (s.a. = 134.0). However, the electrophoretic mobilities of the pools were similar, with most protein bands being concentrated around three main regions, i.e., protein bands with an apparent mr of 100 kDa, of 38-37 kDa and 30 kDa. However, no significant differences were observed in amidolytic activity on the synthetic substrate Tos-Gly-Pro-Arg-pNa, and there was no correlation between thrombin-like and amidolytic activities. A 32 kDa protein endowed with thrombin-like activity and specific activity of 2444 recognized and neutralized by horse anti-B. atrox antivenom, was purified by the successive use of gel filtration, electrofocusing and reverse phase HPLC.
authors.affiliation: Harvard Medical School, Boston, MA, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Cavinato
authors.forename: H
authors.initials: H
authors.lastname: Remold
authors.forename: T L
authors.initials: TL
authors.lastname: Kipnis
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D017837:Bothrops; D001938:Brazil; D002417:Cattle; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006736:Horses; D051379:Mice; D008807:Mice, Inbred BALB C; D012910:Snake Venoms / Q000201:enzymology / Q000302:isolation & purification* / Q000494:pharmacology*
chemical_list: D012910:Snake Venoms