CHARACTERIZATION OF GAMMA-GLUTAMYL-TRANSPEPTIDASE IN THE LIVER OF THE FROG .1. COMPARISON TO THE RAT-LIVER ENZYME

The characteristics of the enzyme gamma-glutamyltranspeptidase were determined in frog liver and compared to those of the rat. In Rana pipiens, tissue distribution studies indicated the order of activity to be: kidney >>> liver >> nerve > egg > lung (similar to) heart > skeletal muscle in homogenates. In the Rana pipiens relative to the Fischer 344 rat, the activity of the liver enzyme was somewhat greater (1.8-fold) and the kidney enzyme substantially less (25-fold). Frog liver gamma-glutamyltranspeptidase displayed strain-dependent differences in activity with Rana pipiens and Rana sylvatica exhibiting comparable activities and Xenopus laevis exhibiting 20-fold lower activities. No influence of sex was apparent in Rana pipiens in contrast to the sex dependent differences observed in the Fischer 344 rat: female : male 4 = 7:1. In homogenates and plasma membrane fractions of Rana pipiens, Xenopus laevis and the Fischer 344 rat, high, and comparable relative specific activities, were observed, 8-11, coupled with protein yields of 2.2-2.5 per cent indicating the enzyme to be plasma membrane bound and associated with the sinusoidal surface of the liver cell. Both the frog Rana pipiens and Xenopus laevis and Fischer 344 rat liver plasma membrane enzymes displayed comparable temperature-induced activation (1.51-1.74-fold) but with a peak for the frogs at 60 degrees C and for the rat at 50 degrees C. Both Acivicin and maleate inhibited the liver plasma membrane gamma-glutamyltranspeptidase of both Rana pipiens and the Fischer 344 rat, but the frog enzyme was less sensitive (89 per cent decrease versus 97 per cent decrease) to 150 mu M Acivicin and more sensitive (65 per cent decrease versus 35 per cent decrease at 150 mM maleate) to maleate. Kinetic studies indicated that the liver plasma membrane enzyme from Rana pipiens had a K-m of 0.61 mM and V-max of 55.6 nmol mg(-1) min(-1) and that from the Fischer 344 rat had a K-m of 3.57 mM and V-max of 71.4 nmol mg(-1) min(-1).