Renal gamma-glutamyltranspeptidase: In situ antiluminal localization

Renal localization of gamma-glutamyltranspeptidase was studied in functioning kidneys perfused with the gamma-glutamyl substrates, gamma-glutamyl-p-nitroanilide and N(gamma-L-glutamyl) 2-naphthylamide. In kidneys perfused with gamma-glutamyl-p-nitroanilide, 90% of the breakdown products appear in the blood (perfusate) rather than urine. The addition of the acceptor substrate glycylglycine to the perfusate stimulated gamma-glutamyl-p-nitroanilide breakdown 3-fold, both on the blood and urine sides, while the appearance of gamma-glutamylglycylglycine confirmed that the activity responsible stems indeed from gamma-glutamyltranspeptidase. Ureter ligation reduced the glomerular filtration rate to less than 10% of the control, while the breakdown of gamma-glutamyl-p-nitroanilide decreased only 5%. Kidneys perfused with N(gamma-L-glutamyl) 2-naphthylamide and the diazonium salt demonstrated staining of the glomeruli; isolated glomeruli and microvessels gave heavy staining when incubated with the histochemical substrates. These results demonstrate the association of a significant portion of the total renal gamma-glutamyltranspeptidase activity with the microvascular compartment.