Comparative studies of kinetic and optical properties of rabbit muscle, sturgeon muscle, and yeast pyruvate kinase.
The kinetic and optical properties of pyruvate kinase isolated from rabbit muscle, sturgeon muscle, and yeast were compared using various activating divalent metal ions as probes for functional features and using ultraviolet circular dichroism (cd) measurements for conformational features, respectively. All three preparations of pyruvate kinase were similar in many aspects, such as activating efficiencies of the four activating metal ions, Mg(II), Co(II), Mn(II), and Ni(II) and pH-rate profiles, suggesting the presence of a similar metal binding locus of these enzymes as well as a common underlying mechanism of action. L-Phe inhibited the rabbit muscle enzyme and turned the hyperbolic kinetics into a sigmoidal kinetic with respect to phosphoenolpyruvate at alkaline pH, while fructose-1,6-biphosphate activated the sturgeon muscle and yeast enzymes and turned the sigmoidal kinetics into hyperbolic kinetics with respect to phosphoenolpyruvate. The ultraviolet cd spectral changes qualitatively correlated well with kinetic observations of all three native enzymes in the presence and absence of allosteric effectors. Our results suggested that there are at least two conformational states of pyruvate kinase which are inducible by the binding of substrate and (or) allosteric effectors. The conformational changes from one form to another in these enzymes are very similar, especially between the rabbit and sturgeon muscle enzymes.
D000818:Animals; D002942:Circular Dichroism; D003035:Cobalt / Q000494:pharmacology; D004789:Enzyme Activation; D005399:Fishes / Q000378:metabolism; D005635:Fructosediphosphates / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009132:Muscles / Q000201:enzymology; D009532:Nickel / Q000494:pharmacology; D010649:Phenylalanine / Q000494:pharmacology; D011770:Pyruvate Kinase / Q000037:antagonists & inhibitors; D011817:Rabbits; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013045:Species Specificity
D005635:Fructosediphosphates; D003035:Cobalt; D008345:Manganese; D010649:Phenylalanine; D009532:Nickel; D011770:Pyruvate Kinase; D008274:Magnesium