Changes in the kinetic parameters of hepatic gamma-glutamyltransferase from streptozotocin-induced diabetic rats.
Previous research has shown that the enzymatic activity of hepatic gamma-glutamyltransferase was increased in streptozotocin-induced diabetic rats with no increase in the expression of the protein. The current work has characterized the differences in the kinetic properties of hepatic gamma-glutamyltransferase from diabetic versus control rats. Hepatic gamma-glutamyltransferase was purified from control male and female rats and from rats made diabetic 30 days previously with streptozotocin. The maximal velocity and the Michaelis constant were determined for the purified enzyme with two separate donors (L-gamma-glutamyl-p-nitroanilide or L-gamma-glutamyl-(7-amido-4-methylcoumarin)) in the presence of one of eight acceptors (L-alanine-glycine, L-glycine-glycine, L-methionine, L-glutamate, L-alanine, L-glutamine, L-phenylalanine or L-aspartate). With both donors, hepatic gamma-glutamyltransferase from diabetic rats had a consistently higher kinetic efficiency than gamma-glutamyltransferase from controls. The kinetic efficiency percent increase of diabetic over control gamma-glutamyltransferase when averaged across all acceptors was higher in males than in females. With L-gamma-glutamyl-p-nitroanilide, the kinetic efficiency increase of diabetic over control gamma-glutamyltransferase was higher with poor acceptors than with highly efficient acceptors. These data indicate that there are differences in the physical properties of hepatic gamma-glutamyltransferase from diabetic versus control rats and from female versus male rats.
Cornwell
P D
PD
Medical Sciences, Indiana University School of Medicine, Jordan Hall 105, Bloomington, IN 47405, USA.
Watkins
J B
JB
D000818:Animals; D003374:Coumarins / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology*; D005260:Female; D005973:Glutamine / Q000031:analogs & derivatives* / Q000378:metabolism; D006031:Glycosylation; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D013311:Streptozocin; D005723:gamma-Glutamyltransferase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D003374:Coumarins; D005973:Glutamine; D013311:Streptozocin; C027245:7-(gamma-glutamyl)-4-methylcoumarylamide; C001300:gamma-glutamine-4-nitroanilide; D005723:gamma-Glutamyltransferase