A new affinity adsorbent for the purification of phospholipases A1 and A2 from animal venoms.
Dimethyl-DL-2,3-distearoyloxy-propyl-2'-hydroxy-ethylammonium++ + (Rosenthal's inhibitor) was coupled to carboxyhexyl-Sepharose 4B, through carbodiimide chemistry. Phospholipase A2 from Heloderma horridum horridum and Crotalus adamanteus bind to the immobilized ligand in the presence of Ca2+ and can be easily eluted under acidic conditions or in the presence of a chelating agent, respectively. This affinity media proved to be effective also in the purification of a Ca2(+)-independent phospholipase A1 from vespid venom.
Vargas-Villarreal
J
J
Centro de Investigación sobre Ingeniería Genética y Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca.
Martin-Polo
J J
JJ
Reynaud
E
E
Alagon
A C
AC
D000818:Animals; D002846:Chromatography, Affinity; D003435:Crotalid Venoms / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D008024:Ligands; D008116:Lizards; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D054466:Phospholipases A1; D054467:Phospholipases A2; D012685:Sepharose; D013228:Stearates; D014688:Venoms / Q000737:chemistry*; D014862:Wasp Venoms / Q000737:chemistry*
D003435:Crotalid Venoms; D008024:Ligands; D013228:Stearates; D014688:Venoms; D014862:Wasp Venoms; C015161:Rosenthal's inhibitor; D012685:Sepharose; D010741:Phospholipases A; D054466:Phospholipases A1; D054467:Phospholipases A2