1,4‐Dihydropyridine receptor associated with Ca2+ channels in human embryonic fibroblasts

FEBS Letters

Dudkin, Sergei M.¹  Chernyuk, Natalia N.¹  Gnedoj, Sergei N.¹  Soldatov, Nikolai M.¹ 

[1] Institute of Biomedical Technology, USSR Ministry of Public Health, 6 Schukinskaya str., 123436 Moscow, USSR

关键词: Dihydropyridine receptor;    Ca2+ channel;    Proliferation;    (Human embryonic fibroblast);    BSA;    bovine serum albumin;    DHP;    1;    4-dihydropyridine;    HEF;    human embryonic fibroblast;    [3H]PMD;    maximum receptor density;    K d;    receptor dissociation constant;   

DOI  :  10.1016/0014-5793(88)80458-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

By using the radioactively labeled 1,4-dihydropyridine (DHP) probe, [³H]PMD, we have demonstrated that cultured human embryonic fibroblasts grown at a low density in Eagle's medium supplemented with serum contain a single class of non-interacting DHP binding sites (B _max, 1.2±0.3 pmol/ 10⁶ cells; K _d, 3.9 nM). After inhibition of the DHP receptor biosynthesis by cycloheximide, the number of [³H]PMD binding sites is reduced with a half-time of 12 h, which implies a turnover rate of 30 000±7500 receptors/h per cell. With progression to confluency, the B _max value decreased up to 0.28±0.08 pmol/10⁶ cells without significant change in K _d value. When cells were grown at a low density in serum-free conditions, the number of [³H]PMD binding sites gradually increased 1.9-fold within 3 days. Addition of serum reversed this effect with the same time course. These results imply that the DHP-sensitive Ca²⁺ channels are involved in the control of the proliferation of human embryonic fibroblasts.

【 授权许可】

Unknown