Fideu M D, Miras-Portugal M T
Departamento de Bioquímica, Facultad de Veterinaria, Universidad Complutense de Madrid, Spain.
Neurochem Res. 1992 Nov;17(11):1099-104. doi: 10.1007/BF00967287.
The adenosine transport in cultured chromaffin cells was increased by the presence of triiodo-L-thyronine (T3) throughout the prolonged period studied. The Vmax values of this transport obtained in absence and presence of 1 microM T3 were 36.21 +/- 2.1 and 44.17 +/- 3.5 (means +/- SD) pmol/10(6) cells/min respectively for 26 hours incubation-time with the hormone. The Km values were not significantly modified. The number of adenosine transporters in cultured chromaffin cells, measured by [3H]nitrobenzylthioinosine (NBTI) binding, was increased by 1 microM T3 for 26 hours incubation-time. The values of binding sites per cell were 33,500 +/- 3,000 and 40,153 +/- 3,700 in absence and presence of T3 respectively, without changing the Kd constant. When the transport studies were carried out in presence of cycloheximide, an inhibitor of protein synthesis, the adenosine transport capacity decreased with a half-life values of 23.9 +/- 2.8 and 24.3 +/- 2.1 hours both in the presence or absence of T3 respectively. When cells were incubated in the presence of both T3 and cycloheximide, not only the activatory effect of T3 was completely abolished but also adenosine transport was decreased to the same extent as with cycloheximide alone. These results indicated that T3 activation of adenosine transport in chromaffin cells required the protein-synthesizing mechanism.
在整个研究的延长期间,三碘 - L - 甲状腺原氨酸(T3)的存在增加了培养的嗜铬细胞中的腺苷转运。在不存在和存在1μM T3的情况下,对于与该激素孵育26小时,这种转运的Vmax值分别为36.21±2.1和44.17±3.5(平均值±标准差)pmol / 10⁶细胞/分钟。Km值没有显著改变。通过[³H]硝基苄硫基肌苷(NBTI)结合测量的培养嗜铬细胞中腺苷转运体的数量,在1μM T3孵育26小时后增加。在不存在和存在T3的情况下,每个细胞的结合位点数分别为33,500±3,000和40,153±3,700,而Kd常数不变。当在蛋白质合成抑制剂环己酰亚胺存在下进行转运研究时,无论是否存在T3,腺苷转运能力均下降,半衰期值分别为23.9±2.8和24.3±2.1小时。当细胞在T3和环己酰亚胺两者存在下孵育时,不仅T3的激活作用完全被消除,而且腺苷转运下降到与单独使用环己酰亚胺相同的程度。这些结果表明,T3对嗜铬细胞中腺苷转运的激活需要蛋白质合成机制。