Rabbit liver phosphofructokinase: rapid purification and phosphorylation site identification.
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.
Zhao
Z
Z
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331, USA.
Pascalar
R W
RW
Malencik
D A
DA
Anderson
S R
SR
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology; D002850:Chromatography, Gel; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D051379:Mice; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010732:Phosphofructokinase-1 / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D010748:Phosphopeptides / Q000737:chemistry / Q000302:isolation & purification; D010766:Phosphorylation; D011479:Protamines; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
D010446:Peptide Fragments; D010748:Phosphopeptides; D011479:Protamines; D000255:Adenosine Triphosphate; D010732:Phosphofructokinase-1