Effect of NAD on flounder muscle glyceraldehyde 3-phosphate dehydrogenase.
Flounder muscle (Pseudopleuronectes americanus) glyceraldehyde-3-phosphate dehydrogenase was characterized as to its stability towards various inactivating treatments in the presence and absence of the enzyme cofactor, NAD. Incubation of a partially purified enzyme preparation at urea concentrations greater than 2 M produced a very rapid inactivation. NAD greatly reduced the rate of inactivation at all the urea concentrations tested. Incubation of each of the three major muscle enzyme forms in 0.1 percent trypsin or chymotrypsin for forty-five minutes decreased the activity of each form by 65 percent and 55 percent, respectively. NAD (5mM) afforded complete protection to each enzyme form from proteolytic digestion by these two enzymes. Exposure of each form to 50 degrees or 20 mM ATP also led to gross inactivation which could be greatly reduced if the respective incubations were performed in the presence of 5mM NAD. NAD was also found to be required for the renaturation of the unfolded urea-denatured subunits to form the active tetramer.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002918:Chymotrypsin; D004355:Drug Stability; D005399:Fishes / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology*; D009243:NAD / Q000494:pharmacology*; D011489:Protein Denaturation; D014357:Trypsin; D014508:Urea
D009243:NAD; D000255:Adenosine Triphosphate; D014508:Urea; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D002918:Chymotrypsin; D014357:Trypsin