Comparative studies of pyruvate kinase from PSE and normal pig muscles.
A fast breakdown of glycogen is observed in muscles of stress-susceptible pigs leading to pale, soft and exudative (PSE) meat. We report a comparative study of pyruvate kinase from muscles of normal and PSE-prone pigs. Compared with the enzyme from normal muscle, pyruvate kinase isolated from PSE muscle shows a five times lower Michaelis constant, Km, for phosphoenol pyruvate and a more than ten times higher Kcat/Km value. The pH dependency of the enzymatic activity is shifted to more acidic values for pyruvate kinase from PSE muscles. According to isoelectric focusing, pyruvate kinase from PSE muscle consists of three isoforms, while only two isoforms are detectable in pyruvate kinase preparations from normal pigs. The various isoforms were isolated by preparative isoelectric focusing and their steady-state properties were compared. Isoform 3, which is found only in PSE muscle, shows a 10-fold higher specific activity, a 30-fold lower Km value and a 100-fold increased kcat/Km value for phosphoenol pyruvate as compared to isoform 1. The presence of isoform 3 in PSE muscle appears to be responsible for the high activity of this enzyme under the more acidic conditions prevailing in PSE muscle. In vitro phosphorylation and dephosphorylation experiments using total enzyme and purified isoenzyme 1 suggest that isoforms 2 and 3 arise from isoform 1 by phosphorylation. Thus protein phosphorylation seems to be responsible for the shift in activity of pyruvate kinase, a key enzyme of glycolysis, under the acidic conditions of PSE muscles.
Schwägele
F
F
Institut für Chemie und Physik, Bundesanstalt für Fleischforschung, Kulmbach, Germany.
Haschke
C
C
Krauss
G
G
Honikel
K O
KO
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008460:Meat; D018482:Muscle, Skeletal / Q000201:enzymology*; D010766:Phosphorylation; D011770:Pyruvate Kinase / Q000302:isolation & purification / Q000378:metabolism*; D013312:Stress, Physiological; D013552:Swine
D007527:Isoenzymes; D011770:Pyruvate Kinase