Synthesis and secretion of an atriopeptin-like protein in rat kidney cell culture.
Synthesis and secretion of an atriopeptin-like protein in rat kidney cell culture.
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大鼠肾细胞培养物中心房肽样蛋白的合成和分泌。
DOI：
10.1172/jci114973
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Greenwald,JE
中科院分区：
文献类型：
--
作者：
Ritter,D;Needleman,P;Greenwald,JE
在培养的新生大鼠肾细胞中，已经证实了心房肽（AP）样激素原（AP 126 ir）的合成和分泌。AP 126 ir可以检测到的细胞提取物和培养的新生和成年大鼠的肾细胞的培养基中使用的酶免疫分析特异性心脏AP激素原。在反相高效液相色谱上，从提取物和培养基中获得的AP与心脏AP原激素共迁移。在培养基中与凝血酶孵育肾AP导致产生一个单一的低分子量峰，其与心脏羧基末端28-氨基酸AP一起迁移。用[35 S]蛋氨酸脉冲的新生肾细胞分泌放射性标记的AP 126 IR，其通过免疫沉淀和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳色谱法检测。孵育的新生儿肾细胞培养物与蛋白质合成抑制剂放线菌酮导致细胞和媒体AP显着减少。用放线菌酮处理新生儿心房培养物时，未检测到细胞和介质AP减少。这些数据证明了新生大鼠肾脏培养物中AP蛋白酶样蛋白的从头合成。此外，与心房不同，肾细胞似乎仅通过组成性方式分泌AP。在原代成年大鼠肾脏培养物中，在皮质小管部分中检测到大部分AP 126 ir，表明这些细胞在成年大鼠肾脏中分泌AP 126 ir。我们假设肾脏AP可能是重要的自分泌或旁分泌调节肾功能。图片
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.Images