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噬菌体T4前头部组装所需基因31蛋白的纯化及特性

Purification and properties of the bacteriophage T4 gene 31 protein required for prehead assembly.

作者信息

Castillo C J, Black L W

出版信息

J Biol Chem. 1978 Apr 10;253(7):2132-9.

PMID:416025
Abstract

A low molecular weight (approximately 16,000), early protein is characterized as the product of the essential T4 head assembly gene 31. This gene is known to be required to allow formation of any ordered head structure from the major T4 capsid protein, P23 (Laemmli, U.K., Beguin, F., and Gujer-Kellenberger, G. (1970) J. Mol. Biol. 47, 69-85). In wild type infection P31 synthesis ceases at late times; in contrast, P31 is overproduced in certain early or regulatory T4 mutant infections, particularly gene 55 mutant infections. P31 was purified preparatively from Escherichia coli infected with the latter mutant, but could only be obtained for the most part in modified form, possibly due to unusual sensitivity to a proteolytic activity. P31 is not cleaved in vivo during normal head assembly, nor does it become a part of the mature head or any ordered prehead structure as determined by an immunological assay using antiserum prepared against the purified protein. However P31 does appear to become a part of the unordered P23 aggregates (lumps) which accumulate when ordered P23 assembly is prevented. We cound find no evidence for P31 association with T4 DNA or the host membrane. Our experiments favor the hypothesis that P31 directly affects the aggregation state and solubility properties of P23.

摘要

一种低分子量(约16,000)的早期蛋白被鉴定为必需的T4头部组装基因31的产物。已知该基因是由主要T4衣壳蛋白P23形成任何有序头部结构所必需的(Laemmli, U.K., Beguin, F., and Gujer-Kellenberger, G. (1970) J. Mol. Biol. 47, 69 - 85)。在野生型感染中,P31的合成在后期停止;相反,在某些早期或调节性T4突变体感染中,特别是基因55突变体感染中,P31会过量产生。P31是从感染了后一种突变体的大肠杆菌中通过制备性方法纯化得到的,但大部分只能以修饰形式获得,这可能是由于对蛋白水解活性异常敏感所致。在正常头部组装过程中,P31在体内不会被切割,通过使用针对纯化蛋白制备的抗血清进行免疫测定确定,它也不会成为成熟头部或任何有序前头部结构的一部分。然而,当有序的P23组装被阻止时,P31似乎确实成为了无序的P23聚集体(块状物)的一部分。我们没有发现P31与T4 DNA或宿主膜相关的证据。我们的实验支持这样一种假设,即P31直接影响P23的聚集状态和溶解性。

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