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磷脂修饰会延缓小鼠MOPC-31c浆细胞瘤细胞对免疫球蛋白G1的细胞内转运和分泌。

Phospholipid modification retards intracellular transport and secretion of immunoglobulin G1 by mouse MOPC-31c plasmacytoma cells.

作者信息

Nakano A, Maeda M, Nishijima M, Akamatsu Y

出版信息

Biochim Biophys Acta. 1982 Nov 8;692(2):177-86. doi: 10.1016/0005-2736(82)90519-3.

Abstract

The intracellular transport and secretion of immunoglobulin G1(IgG1) by mouse MOPC-31C plasmacytoma cells were analyzed from the viewpoint of the roles of phospholipids. The membrane phospholipids were modified by culturing cells in a medium supplemented with choline analogues, N,N'-dimethylethanolamine or N-monomethylethanolamine, and accordingly the membranes were enriched in phosphatidyl-N,N'-dimethylethanolamine or phosphatidyl-N-monomethylethanolamine (Maeda, M., Tanaka, Y. and Akamatsu, Y. (1980) Biochem. Biophys. Res. Commun. 96, 876-881). The modified cells were pulse-labeled with L-[35S]methionine and the secretion of labeled IgG1 was chased. Half of the IgG1 was exported to the extracellular medium 1-1.5 h and 2-3 h after synthesis by choline- and dimethylethanolamine-supplemented cells, respectively. However, most of the newly synthesized IgG1 was not secreted by monomethylethanolamine-supplemented cells, even after 5 h; it remained within the cells. The sensitivity of intracellular IgG1 to endoglycosidase H was examined for probing the movement of IgG1 from the rough endoplasmic reticulum to the Golgi complex. Half of the newly synthesized IgG1 acquired resistance to endoglycosidase H after 30-45 min, 1-1.5 h and 2-3 h in choline-, dimethylethanolamine- and monomethylethanolamine-supplemented cells, respectively. Thus, the transport of IgG1 was markedly retarded by the modification with choline analogues, dimethylethanolamine or monomethylethanolamine, at least in the following two processes, from the rough endoplasmic reticulum to the Golgi complex and from the Golgi to the outside of cells. Modification with monomethylethanolamine was more effective than that with dimethylethanolamine in slowing down the transport of IgG1 and appeared to cause accumulation of IgG1 within the cells. A morphological study was also carried out for the three kinds of cell. The roles of phospholipids in the processes of membrane flow are discussed.

摘要

从小鼠MOPC - 31C浆细胞瘤细胞中免疫球蛋白G1(IgG1)的细胞内运输和分泌角度,对磷脂的作用进行了分析。通过在补充胆碱类似物、N,N'-二甲基乙醇胺或N - 单甲基乙醇胺的培养基中培养细胞来修饰膜磷脂,相应地,膜中富含磷脂酰 - N,N'-二甲基乙醇胺或磷脂酰 - N - 单甲基乙醇胺(前田真、田中洋和赤松洋(1980年)《生物化学与生物物理研究通讯》96卷,第876 - 881页)。用L - [35S]甲硫氨酸对修饰后的细胞进行脉冲标记,并追踪标记的IgG1的分泌情况。分别在补充胆碱和二甲基乙醇胺的细胞合成IgG1后1 - 1.5小时和2 - 3小时,一半的IgG1被分泌到细胞外培养基中。然而,即使在5小时后,补充单甲基乙醇胺的细胞中大多数新合成的IgG1也未分泌;它仍留在细胞内。通过检测细胞内IgG1对内切糖苷酶H的敏感性,来探究IgG1从糙面内质网到高尔基体复合体的转运情况。在分别补充胆碱、二甲基乙醇胺和单甲基乙醇胺的细胞中,新合成的IgG1分别在30 - 45分钟、1 - 1.5小时和2 - 3小时后获得了对内切糖苷酶H的抗性。因此,至少在从糙面内质网到高尔基体复合体以及从高尔基体到细胞外这两个过程中,用胆碱类似物、二甲基乙醇胺或单甲基乙醇胺修饰会显著阻碍IgG1的转运。用单甲基乙醇胺修饰比用二甲基乙醇胺修饰更能有效地减缓IgG1的转运,并且似乎导致IgG1在细胞内积累。还对这三种细胞进行了形态学研究。讨论了磷脂在膜流动过程中的作用。

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