title: Purification of homogeneous rat phosphofructokinase isozymes with high specific activities.
abstract: The purification of rat muscle and liver phosphofructokinase (PFK) isozymes has been greatly facilitated by column chromatographic separation on immobilized Cibacron Blue F3GA. The homogeneous liver PFK isozyme exhibited a specific activity of greater than 200 units per mg of protein which is nearly two-fold greater than has been previously reported for this isozyme. The yields for this isozyme exceeded 40% of the original activity and the molecular weight of its subunit was about 85,000 as determined by SDS-polyacrylamide gel electrophoresis. The muscle PFK isozyme's specific activity was approximately 265 units/mg of protein which also is about twice the greatest specific activity previously reported. The overall yield for muscle PFK exceeded 50% of the original activity, and the molecular weight of its subunit was approximately 82,000. Using each homogeneous isozyme, antibodies were produced in rabbits; and the immunoglobin-G (IgG) fraction from the sera of these rabbits was highly specific for the PFK isozyme used as an antigen.
mesh_terms: D000818:Animals; D003429:Cross Reactions; D007527:Isoenzymes / Q000302:isolation & purification*; D008099:Liver / Q000201:enzymology; D008297:Male; D009132:Muscles / Q000201:enzymology; D010732:Phosphofructokinase-1 / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D007527:Isoenzymes; D010732:Phosphofructokinase-1