Isolation of membrane-bound renal enzymes that metabolize kinins and angiotensins.
Cortex of rat kidney was homogenized and fractions enriched in plasma membrane, endoplasmic reticulum or brush border were prepared by several techniques of differential centrifugation. The identity and homogeneity of the membrane fragments were investigated by assaying marker enzymes and by transmission and scanning electron microscopy. Kallikrein was present in both plasma-membrane- and endoplasmic-reticulum-enriched fractions isolated by two fractionation procedures. Kallikrein was highly concentrated in a plasma-membrane fraction but was absent from the brush-border membrane of proximal tubular cells. Cells of transplanted renal tumours of the rat, originating from the proximal tubule, had no kallikrein activity. Kininase activity, angiotensin I-converting enzyme (kininase II) and angiotensinase were found in a plasma-membrane-enriched fraction and especially in the fraction containing isolated brush border. It is suggested that after renal kallikrein is synthesized on endoplasmic reticulum, it is subsequently reoriented to a surface membrane for activation and release. Renal kallikrein may enter the tubular filtrate distal to the proximal tubules. The brush-border membrane of proximal tubule is the major site of inactivation of kinins and angiotensin II..
D000251:Adenosine Triphosphatases / Q000032:analysis; D000469:Alkaline Phosphatase / Q000032:analysis; D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology; D010450:Endopeptidases / Q000032:analysis; D004721:Endoplasmic Reticulum / Q000201:enzymology; D005952:Glucose-6-Phosphatase / Q000032:analysis; D007610:Kallikreins / Q000032:analysis; D007672:Kidney Cortex / Q000201:enzymology / Q000648:ultrastructure; D007680:Kidney Neoplasms / Q000201:enzymology; D007687:Kidney Tubules, Proximal / Q000201:enzymology*; D007705:Kinins / Q000378:metabolism*; D001121:Lysine Carboxypeptidase / Q000032:analysis; D008297:Male; D008861:Microsomes / Q000201:enzymology; D007703:Peptidyl-Dipeptidase A / Q000032:analysis; D051381:Rats
D007705:Kinins; D000804:Angiotensin II; D000469:Alkaline Phosphatase; D005952:Glucose-6-Phosphatase; D010450:Endopeptidases; D007703:Peptidyl-Dipeptidase A; D001121:Lysine Carboxypeptidase; D007610:Kallikreins; D000251:Adenosine Triphosphatases