INTERACTION OF TUBULIN WITH DRUGS AND ALKYLATING-AGENTS .1. ALKYLATION OF TUBULIN BY IODO-ACETAMIDE-C-14 AND N,N'-ETHYLENEBIS(IODOACETAMIDE)
INTERACTION OF TUBULIN WITH DRUGS AND ALKYLATING-AGENTS .1. ALKYLATION OF TUBULIN BY IODO-ACETAMIDE-C-14 AND N,N'-ETHYLENEBIS(IODOACETAMIDE)
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DOI：
10.1021/bi00518a031
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发表时间：
1981-01-01
期刊：
影响因子：
2.9
通讯作者：
ROACH, MC
中科院分区：
文献类型：
--
作者：
LUDUENA, RF;ROACH, MC
据报道，[牛脑]微管蛋白的SH基团在调节微管组装和秋水仙碱与微管蛋白的结合中起作用。用烷基化剂碘[14C]乙酰胺及其双功能类似物N，N''-乙烯双（碘乙酰胺）作为微管蛋白SH基团的探针。在8 M尿素存在下，。-和。beta。-微管蛋白分别具有10-11和8烷基化SH，其中一个高分子量蛋白（HMW 2）具有5 SH/MW 271,000。在没有尿素的情况下，. α的烷基化速率。-和。beta。-微管蛋白明显降低，但HMW 2不受影响。微管蛋白的SH被完整的微管所掩盖。N，N′-乙二（碘乙酰胺）与游离微管蛋白反应生成一个带，命名为。β。*，它在.beta之前迁移。聚丙烯酰胺凝胶。*似乎表示。beta的一种形式。-微管蛋白在SH基团之间至少含有1个链内交联。形成。*在完整的微管中被抑制，如果微管蛋白被8 M尿素，1%十二烷基硫酸钠或煮沸变性，则被废除。因此，N，N''-乙二（碘乙酰胺）可用作游离微管蛋白天然构象的探针。
The SH groups of [bovine brain] tubulin are reported to play a role in regulating microtubule assembly and colchicine binding to tubulin. The alkylating agents iodo[14C]acetamide and its bifunctional analogue N,N''-ethylenebis(iodacetamide) were used as probes for the SH groups of tubulin. In the presence of 8 M urea, .alpha.- and .beta.-tubulin have 10-11 and 8 alkylatable SH, respectively, and one of the high MW proteins (HMW 2) has 5 SH/MW 271,000. In the absence of urea, the rates of alkylation of .alpha.- and .beta.-tubulin are significantly lower but that of HMW 2 is unaffected. The SH of tubulin were masked in intact microtubules. N,N''-Ethylenebis (iodoacetamide) reacts with free tubulin to generate a band, designated .beta.*, which migrates ahead of .beta. on polyacrylamide gels. .beta.* appears to represent a form of .beta.-tubulin containing at least 1 intrachain cross-link between SH groups. Formation of .beta.* is inhibited in intact microtubules and is abolished if tubulin is denatured by 8 M urea, 1% sodium dodecyl sulfate, or boiling. N,N''-Ethylenebis (iodoacetamide) may thus be used as a probe for the native conformation of free tubulin.