SELECTIVE CARBETHOXYLATION OF HISTIDINE RESIDUES OF ACTIN BY DIETHYLPYROCARBONATE
SELECTIVE CARBETHOXYLATION OF HISTIDINE RESIDUES OF ACTIN BY DIETHYLPYROCARBONATE
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DOI：
10.1111/j.1432-1033.1974.tb03452.x
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发表时间：
1974-01-01
期刊：
影响因子：
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通讯作者：
MUHLRAD, A
中科院分区：
文献类型：
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作者：
HEGYI, G;PREMECZ, G;MUHLRAD, A
肌动蛋白的组氨酸残基与焦碳酸二乙酯carbethoxylated。四个和三个组氨酸残基每个肌动蛋白单体迅速carbethoxylated在G和F的形式，分别。在G-肌动蛋白的部分羧乙氧羰基化后，可以通过超离心分离不可聚合的部分。对这些组分的分析表明，G-肌动蛋白的四种快速反应组氨酸之一在聚合中起着至关重要的作用。通过羟胺处理去除组氨酸残基上的乙氧羰基，恢复了聚合的能力。使用[14 C]测试了反应的特异性。在胰蛋白酶消化不可聚合的乙氧羰基化肌动蛋白后，通过凝胶过滤分离单个组氨酰肽，对角电泳聚合所必需的组氨酸残基被鉴定为组氨酸-40。
Histidine residues of actin were carbethoxylated with diethylpyrocarbonate. Four and three histidine residues per actin monomer were rapidly carbethoxylated in the G and F form, respectively. After partial Carbethoxylation of G‐actin a non‐polymerizable fraction could be separated by ultracentrifugation. The analysis of these fractions showed that one of the four fast‐reacting histidines of G‐actin plays an essential role in polymerization.Removal of the carbethoxy groups from the histidine residues by hydroxylamine treatment restored the ability to polymerize.The specificity of the reaction was tested by the use of [14C]diethylpyrocarbonate.After tryptic digestion of non‐polymerizable carbethoxylated actin a single histidyl peptide was isolated by gel filtration and diagonal electrophoresis. The histidine residue essential for polymerization was identified as histidine‐40.