培养的豚鼠巨核细胞对肌动蛋白的合成。与纤维蛋白的复合物形成。

Synthesis of actin by cultured guinea pig megakaryocytes. Complex formation with fibrin.

作者信息

Nachman R, Levine R, Jaffe E

出版信息

Biochim Biophys Acta. 1978 Sep 21;543(1):91-105. doi: 10.1016/0304-4165(78)90457-9.

Abstract

Guinea pig megakaryocytes were isolated from femoral marrow and cultured in the presence of radioactive amino acids. Radioactivity was incorporated into several proteins including a 42 000 dalton polypeptide identified as actin by DNAase agarose affinity chromatography. Quantitative immunoelectrophoresis of megakaryocyte extract revealed that 3.0% of the total solubilized cellular protein was fibrinogen. Immunoabsorption studies using anti guinea pig fibrinogen beads failed to reveal the presence of newly synthesized radioactive fibrinogen in the cellular extract, however, radioactive actin was detected in the eluates obtained from the immune beads. When guinea pig fibrinogen was clotted with thrombin in the presence of radioactive megakaryocyte extract, a complex formed between a high molecular weight species of fibrin and actin. No actin fibrinogen complex was detected. The results suggest that actin synthesized by megakaryocytes complexes with fibrin formed from a relatively large pool of non-radioactive intracellular fibrinogen.

摘要

豚鼠巨核细胞从股骨骨髓中分离出来，并在放射性氨基酸存在的情况下进行培养。放射性被掺入到几种蛋白质中，包括一种通过DNA酶琼脂糖亲和层析鉴定为肌动蛋白的42000道尔顿多肽。巨核细胞提取物的定量免疫电泳显示，总可溶性细胞蛋白的3.0%是纤维蛋白原。使用抗豚鼠纤维蛋白原珠的免疫吸收研究未能揭示细胞提取物中新合成的放射性纤维蛋白原的存在，然而，在从免疫珠获得的洗脱液中检测到了放射性肌动蛋白。当在放射性巨核细胞提取物存在的情况下，用凝血酶使豚鼠纤维蛋白原凝结时，一种高分子量的纤维蛋白与肌动蛋白之间形成了复合物。未检测到肌动蛋白纤维蛋白原复合物。结果表明，巨核细胞合成的肌动蛋白与由相对大量的非放射性细胞内纤维蛋白原形成的纤维蛋白结合。