INCORPORATION OF L-TYROSINE, L-PHENYLALANINE AND L-3,4-DIHYDROXYPHENYLALANINE AS SINGLE UNITS INTO RAT-BRAIN TUBULIN
INCORPORATION OF L-TYROSINE, L-PHENYLALANINE AND L-3,4-DIHYDROXYPHENYLALANINE AS SINGLE UNITS INTO RAT-BRAIN TUBULIN
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DOI：
10.1111/j.1432-1033.1975.tb02435.x
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发表时间：
1975-01-01
期刊：
影响因子：
--
通讯作者：
CAPUTTO, R
中科院分区：
文献类型：
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作者：
ARCE, CA;RODRIGUEZ, JA;CAPUTTO, R
将[14 C]酪氨酸作为单个单元掺入到大鼠脑匀浆的可溶性级分的蛋白质中的产物通过以下用于纯化微管蛋白的程序来纯化。纯化材料的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示含有所有放射性的单一蛋白质条带。纯化数据表明，该蛋白质占上清液级分总蛋白质的10.2%。这与通过[3 H]秋水仙碱结合方法发现的微管蛋白的量（总蛋白的10.5%）非常一致。在微管蛋白的α亚基中发现掺入的[14 C]-酪氨酸。用硫酸长春碱沉淀[3 H]秋水仙碱和[14 C]酪氨酸标记的蛋白质，沉淀物中定量回收了3 H和14 C的放射性（98%）。长春碱沉淀物的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示14 C放射性随微管蛋白带移动。苯丙氨酸和3，4-二羟基苯丙氨酸的实验结果与酪氨酸的实验结果相同。秋水仙碱的结合不干扰酪氨酸的掺入。大约30%的大鼠脑上清液中的微管蛋白可以将酪氨酸作为单一单位掺入。
The product of the incorporation of [14C]tyrosine as single unit into a protein of the soluble fraction of rat brain homogenate was purified by following a procedure used to purify tubulin. Sodium dodecylsulphate‐polyacrylamide gel electrophoresis of the purified material showed a single protein band containing all the radioactivity. Purification data indicate that this protein accounts for 10.2% of the total protein of the supernatant fraction. This is in good agreement with the amount found for tubulin by the [3H]colchicine‐binding method (10.5% of the total protein). The incorporated [14C]‐tyrosine was found in the α‐subunit of tubulin.Protein labelled with [3H]colchicine and [14C]tyrosine was precipitated with vinblastine sulphate and the radioactivity of3H and that of14C were quantitatively recovered in the precipitate (98%). Sodium dodecylsulphate ‐ polyacrylamide gel electrophoresis of the vinblastine precipitate showed that the14C radioactivity moved with the tubulin band.Results obtained in experiments with phenylalanine and 3,4‐dihydroxyphenylalanine were identical to those obtained for tyrosine.Binding of colchicine did not interfere with the incorporation of tyrosine.About 30% of tubulin from rat brain supernatant fraction can incorporate tyrosine as single unit.