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昆虫锥虫目寄生虫——fasiculata短膜虫的组蛋白。

The histones of the insect trypanosomatid, Crithidia fasciculata.

作者信息

Duschak V G, Cazzulo J J

机构信息

Instituto de Investigaciones Bioquímicas Fundación Campomar, Universidad de Buenos Aires, CONICET, Argentina.

出版信息

Biochim Biophys Acta. 1990 Sep 3;1040(2):159-66. doi: 10.1016/0167-4838(90)90071-m.

Abstract

The histone-like proteins of the monogenetic parasite Crithidia fasciculata were extracted with 0.2 M sulfuric acid either from purified nuclei, or from purified chromatin, in both cases in the presence of 1 mM tosyl lysylchloromethylketone and 2 mM phenyl methyl sulfonyl fluoride as proteinase inhibitors. The presence of histones in the flagellate, nonidentical with those from calf thymus used as controls, was shown by their electrophoretic patterns in three different polyacrylamide gel systems; their staining with Alkaline fast green, specific for basic proteins; their global amino acid composition and absorption spectrum and their molecular weights. The protein showing the slower mobility in SDS gels and the fastest mobility in the urea-acetic acid-Triton gels, seems to be an H1 histone, because of its metachromatic staining with Coomassie brilliant blue, solubility characteristics, differential destaining properties and amino acid composition. Band 5 in Triton-urea-acetic acid gels is probably an HMG protein. We conclude that C. fasciculata has a complete set of histones and that the lack of chromosome condensation during mitosis is not due to lack of histone H1.

摘要

单细胞寄生虫纤细短膜虫的类组蛋白可通过0.2M硫酸从纯化的细胞核或纯化的染色质中提取,两种情况下均存在1mM甲苯磺酰赖氨酰氯甲基酮和2mM苯甲基磺酰氟作为蛋白酶抑制剂。通过在三种不同的聚丙烯酰胺凝胶系统中的电泳图谱、用对碱性蛋白特异的碱性固绿染色、整体氨基酸组成和吸收光谱以及分子量,证明了鞭毛虫中存在与用作对照的小牛胸腺组蛋白不同的组蛋白。在SDS凝胶中迁移较慢而在尿素-乙酸- Triton凝胶中迁移最快的蛋白质,由于其用考马斯亮蓝的异染性染色、溶解性特征、差异脱色特性和氨基酸组成,似乎是一种H1组蛋白。Triton-尿素-乙酸凝胶中的条带5可能是一种高迁移率族蛋白。我们得出结论,纤细短膜虫有一整套组蛋白,并且有丝分裂期间缺乏染色体浓缩并非由于缺乏组蛋白H1。

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