Synthesis and secretion of an atriopeptin-like protein in rat kidney cell culture.
The synthesis and secretion of an atriopeptin(AP)-like prohormone (AP126ir) has been demonstrated in rat neonatal renal cell cultures. AP126ir could be detected in the cellular extract and the medium from cultured kidney cells of neonatal and adult rats using an enzyme immunoassay specific for cardiac AP prohormone. On reverse-phase high-performance liquid chromatography, the AP obtained from the extract and the medium comigrated with cardiac AP prohormone. Incubation of the renal AP in the medium with thrombin resulted in the generation of a single low molecular mass peak which migrated with the cardiac carboxy-terminal 28-amino acid AP. Neonatal kidney cells pulsed with [35S]methionine secreted radiolabeled AP126ir, which was detected by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis chromatography. Incubation of neonatal kidney cell cultures with the protein synthesis inhibitor cycloheximide resulted in a significant decrease in both the cellular and media AP. No decrease in cellular and media AP was detected when neonatal atrial cultures were treated with cycloheximide. These data demonstrate the de novo synthesis of an AP prohormone-like protein in neonatal rat kidney cultures. Furthermore, unlike the atria, kidney cells appear to secrete AP solely by constitutive means. In primary adult rat kidney cultures, most of AP126ir was detected in the cortical tubule fraction demonstrating that these cells secrete AP126ir in the adult rat kidney. We hypothesize that the renal AP may be important as an autocrine or paracrine regulator of renal function.
Ritter
D
D
Department of Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63108.
Needleman
P
P
Greenwald
J E
JE
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D009320:Atrial Natriuretic Factor; D003513:Cycloheximide