Aldosterone-induced proteins: purification and localization of GP65,70.
Aldosterone stimulates sodium transport in responsive epithelia by inducing "effector" proteins that control or modulate transcellular sodium flux. We have previously identified a group of electrophoretically microheterogeneous (pI 5.8-6.2) and polymorphic (Mr 65 and 70) glycoproteins that are specifically induced by aldosterone in toad urinary bladders (TUBs) and cultured toad kidney cells (A6 cell line). We raised a series of monoclonal antibodies (MAb) to these proteins and, using light and electron immunohistochemistry, localized the higher Mr glycoproteins (GP70) to the apical plasma membrane and subapical granules of the sodium-transporting cell of the TUB epithelium, the granular cell. GP70 appears to be discharged into the bladder lumen; this process is increased by phorbol myristate acetate, an agent known to induce granule exocytosis. These findings are consistent with the possibility that GP70 represent components or modulators of the "high-resistance" renal epithelial sodium channel. MAbs reactive against GP65 did not identify these glycoproteins within TUB epithelial cells; these lower Mr aldosterone-induced proteins may be incompletely processed forms of GP70.
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H M
HM
Renal Electrolyte Section, Veterans Administration Medical Center, Philadelphia, Pennsylvania.
Weisberg
L
L
Clayman
M
M
Neilson
E
E
Wade
J B
JB
Cox
M
M
D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal; D002024:Bufo marinus; D002462:Cell Membrane / Q000032:analysis; D003594:Cytoplasmic Granules / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000032:analysis* / Q000096:biosynthesis; D006651:Histocytochemistry; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D007668:Kidney / Q000032:analysis* / Q000187:drug effects / Q000648:ultrastructure; D008854:Microscopy, Electron; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D001743:Urinary Bladder / Q000032:analysis* / Q000187:drug effects / Q000648:ultrastructure; D014909:Wheat Germ Agglutinins
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D014909:Wheat Germ Agglutinins; D000450:Aldosterone; D013755:Tetradecanoylphorbol Acetate