Turnover and storage of newly synthesized adenine nucleotides in bovine adrenal medullary cell cultures.
Turnover and storage of newly synthesized adenine nucleotides in bovine adrenal medullary cell cultures.
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牛肾上腺髓质细胞培养物中新合成的腺嘌呤核苷酸的周转和储存。
DOI：
10.1111/j.1471-4159.1986.tb12938.x
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发表时间：
1986
影响因子：
4.7
通讯作者：
Kirshner,N
中科院分区：
文献类型：
--
作者：
Corcoran,JJ;Wilson,SP;Kirshner,N
用32piand [3H]腺苷孵育培养的肾上腺髓细胞，对其腺嘌呤核苷酸储存进行放射性标记，并检测核苷酸的周转、亚细胞分布和分泌情况。ATP占标记腺嘌呤核苷酸的84-88%，ADP占11-13%，AMP占1-3%。32p -腺嘌呤核苷酸和3h -核苷酸的转换是双相的，几乎相同；初始快速期为3.5-4.5 h，半衰期为7 - 17天，取决于特定的细胞制备。标记腺嘌呤核苷酸的慢相与标记儿茶酚胺的慢相相同。标记的腺嘌呤核苷酸的亚细胞分布提供了证据，证明至少有两个腺嘌呤核苷酸池组成了半衰期长的成分。一个包含大部分内源性核苷酸（占总量的75%）的池存在于染色质囊泡中；第二池的亚细胞定位尚未确定。研究还表明，[3H]ATP和[32P]ATP在细胞内的分布不同；标记3天后，75%的[32P]ATP存在于染色质囊泡中，而只有35%的[3H]ATP存在于染色质囊泡中。从分泌物研究中也获得了两种具有长半衰期的ATP池以及[32P]ATP和[3H]ATP差异分布的证据。用[3H]腺苷和32pii标记24小时后，用尼古丁或蝎子毒液刺激细胞培养物释放的儿茶酚胺相对于32p标记的化合物多两倍，相对于3H标记的化合物多三倍。
The adenine nucleotide stores of cultured adrenal medullary cells were radiolabeled by incubating the cells with32Piand [3H]adenosine and the turnover, subcellular distribution, and secretion of the nucleotides were examined. ATP represented 84–88% of the labeled adenine nucleotides, ADP 11–13%, and AMP 1–3%. The turnover of32P‐adenine nucleotides and3H‐nucleotides was biphasic and virtually identical; there was an initial fast phase with a t½ of 3.5–4.5 h and a slow phase with a half‐life varying from 7 to 17 days, depending upon the particular cell preparation. The t½ of the slow phase for labeled adenine nucleotides was the same as that for the turnover of labeled catecholamines. The subcellular distribution of labeled adenine nucleotides provides evidence that there are at least two pools of adenine nucleotides which make up the component with the long half‐life. One pool, which contains the bulk of endogenous nucleotides (75% of the total), is present within the chromaffin vesicles; the subcellular localization of the second pool has not been identified. The studies also show that [3H]ATP and [32P]ATP are distributed differently within the cell; 3 days after labeling 75% of the [32P]ATP was present in chromaffin vesicles while only 35% of the [3H]ATP was present in chromaffin vesicles. Evidence for two pools of ATP with long half‐lives and for the differential distribution of [32P]ATP and [3H]ATP was also obtained from secretion studies. Stimulation of cell cultures with nicotine or scorpion venom 24 h after labeling with [3H]adenosine and32Pireleased relatively twice as much catecholamine as32P‐labeled compounds and relatively three times as much catecholamine as3H‐labeled compounds.