Isoelectric focusing in immobilized pH gradients of a snake venom fibrinolytic enzyme.
A fibrinolytic enzyme with a molecular weight between 23,000 and 25,000 Da has been purified from southern copperhead snake venom. Immobilized pH gradient isoelectric focusing with an ultranarrow pH interval (pH 6.65-6.95) resolved two isoforms of the fibrinolytic enzyme that were not resolved by standard isoelectric focusing. Attempts at purification of the individual isoenzymes by semi-preparative scale IPG and elution of enzyme by macerating the gel yielded only 20-40% recovery of activity. In attempts to improve recovery, a semi-preparative IPG canal-isoelectric focusing technique has been utilized.
Guan
A L
AL
Department of Biochemistry, USC School of Medicine, Los Angeles 90033.
Markland
F S
FS
D003435:Crotalid Venoms / Q000032:analysis*; D005342:Fibrinolysis; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods*; D007527:Isoenzymes / Q000302:isolation & purification
D003435:Crotalid Venoms; D007527:Isoenzymes