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  • Title: Rabbit liver phosphofructokinase: rapid purification and phosphorylation site identification.
    Author: Zhao Z, Pascalar RW, Malencik DA, Anderson SR.
    Journal: Biochem Biophys Res Commun; 1996 May 15; 222(2):410-5. PubMed ID: 8670219.
    Abstract:
    Liver phosphofructokinase can be selectively precipitated by the addition of protamine sulfate to a heat-treated crude extract and redissolved, giving nearly full recovery of catalytic activity in combination with a 67-fold increase in specific activity. We have incorporated protamine sulfate precipitation into a five step purification procedure that can be completed in one day--giving 47% recovery of electrophoretically homogeneous liver phosphofructokinase having a specific activity of 50 units/mg. The radio-labeled fragment isolated from a CNBr digest of liver phosphofructokinase that has undergone in vitro phosphorylation catalyzed by the cAMP-dependent protein kinase has the sequence AEYVSGELEHVTRRSLS.
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