title: Immobilized flounder muscle glyceraldehyde 3-phosphate dehydrogenase.
abstract: Partially purified flounder muscle (Pseudopleuronectus americanus) glyceraldehyde 3-phosphate dehydrogenase was immobilized on cyanogen bromide-activated Sepharose. The catalytic properties of the immobilized preparation were studied to determine if immobilization alters the kinetic properties of the native holoenzyme. The results indicate that the pH activity profile of immobilized glyceraldehyde 3-phosphate dehydrogenase did not differ from that of the native enzyme. The Michaelis constants (Km) for NAD and glyceraldehyde 3-phosphate were somewhat altered. The enzyme stability toward various inactivation treatments in the presence and absence of NAD was characterized and compared to that of he native enzyme. When either form of the enzyme was incubated with urea at concentrations greater than 2M, inactivation occurred very rapidly. Incubation in 0.1% trypsin for 60 minutes decreased the activity of immobilized glyceraldehyde 3-phosphate dehydrogenase by 45% and of the native soluble enzyme by 70%. The immobilized enzyme also exhibited considerably more stability than the native soluble enzyme when exposed to a temperature of 50 degrees or to 20 mM ATP. In all cases NAD either greatly reduced the rate of inactivation or completely protected the enzyme from inactivation.
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002384:Catalysis; D004355:Drug Stability; D004800:Enzymes, Immobilized / Q000378:metabolism; D005399:Fishes; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors / Q000378:metabolism; D006358:Hot Temperature; D008722:Methods; D009132:Muscles / Q000201:enzymology*; D011489:Protein Denaturation; D014508:Urea / Q000494:pharmacology
chemical_list: D004800:Enzymes, Immobilized; D000255:Adenosine Triphosphate; D014508:Urea; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases