Proteins of the kidney microvillar membrane. Effects of monensin, vinblastine, swainsonine and glucosamine on the processing and assembly of endopeptidase-24.11 and dipeptidyl peptidase IV in pig kidney slices.
The effects of various inhibitors were studied on the biogenesis of endopeptidase-24.11 (EC 3.4.24.11) and dipeptidyl peptidase IV (EC 3.4.14.5) in slices of renal cortex, from piglets of the Yucatan strain, maintained in organ culture. These microvillar peptidases were synthesized within membrane compartments and underwent glycosylation to yield high-mannose and complex forms [the preceding paper, Stewart & Kenny (1984) Biochem. J. 224, 549-558]. Monensin caused very gross ultrastructural changes in the proximal-tubular cells, resulting from distension of the Golgi sacs. It blocked the processing of the high-mannose to the complex glycosylated forms of the peptidases and prevented their assembly in the microvillar membrane. Swainsonine, an inhibitor of alpha-mannosidase II, generated new 'hybrid' forms of the proteins, intermediate in Mr between the high-mannose and the complex forms, but did not prevent assembly of the hybrid forms in microvilli. Vinblastine, an agent that affects microtubules, delayed, but did not abolish, either the processing or the transport to microvilli. Glucosamine interfered with the initial glycosylation reactions and generated heterogeneous sets of partially glycosylated polypeptides of lower Mr than the high-mannose forms. These results are discussed in relation to the site and mechanism of glycosylation and the involvement of the Golgi complex and microtubules in the biogenesis of these membrane peptidases.
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D005944:Glucosamine / Q000494:pharmacology; D007672:Kidney Cortex / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008871:Microvilli / Q000201:enzymology; D008985:Monensin / Q000494:pharmacology; D009924:Organ Culture Techniques; D013347:Subcellular Fractions / Q000201:enzymology; D017026:Swainsonine; D013552:Swine; D014747:Vinblastine / Q000494:pharmacology
D000470:Alkaloids; D014747:Vinblastine; D008985:Monensin; D005944:Glucosamine; D017026:Swainsonine