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将一种病毒粒子蛋白激酶鉴定为病毒特异性酶。

Characterization of a virion protein kinase as a virus-specified enzyme.

作者信息

Silberstein H, August J T

出版信息

J Biol Chem. 1976 May 25;251(10):3185-90.

PMID:178663
Abstract

Antibodies which completely inhibited the enzymatic activity of the protein kinase associated with virions of frog virus were obtained by immunization of rabbits with the purified enzyme. This inhibition provided a specific probe for the frog virus protein kinase, since this antiserum had no inhibitory effect on a variety of other protein kinases, including the activity of uninfected cells, or the protein kinase associated with vesicular stomatitis virus or vaccinia virus cultivated in the same cell line as frog virus. The frog virus protein kinase was characterized as a virus-specified protein on the basis of the following observations: (a) the virion protein kinase was antigenically distinct from essentially all of the protein kinase expressed in uninfected cells; (b) following infection by frog virus more than a 15-fold increase was detected in the specific activity of intracellular protein kinase and most of this activity was antigenically related to the virion enzyme; (c) when frog virus was grown in cells derived from widely different species, the antigenic and biochemical specificities of the virion protein kinase remained identical; and (d) screening of cells infected with different temperature-sensitive mutants of frog virus indicated that certain viral mutants failed to synthesize this protein kinase when cultivated at the nonpermissive temperature.

摘要

用纯化的酶免疫兔子,获得了能完全抑制与蛙病毒粒子相关的蛋白激酶酶活性的抗体。这种抑制作用为蛙病毒蛋白激酶提供了一种特异性探针,因为这种抗血清对多种其他蛋白激酶没有抑制作用,包括未感染细胞的活性,或与在与蛙病毒相同细胞系中培养的水疱性口炎病毒或痘苗病毒相关的蛋白激酶。基于以下观察结果,蛙病毒蛋白激酶被鉴定为一种病毒特异性蛋白:(a) 病毒粒子蛋白激酶在抗原性上与未感染细胞中表达的基本上所有蛋白激酶都不同;(b) 蛙病毒感染后,细胞内蛋白激酶的比活性增加了15倍以上,且大部分活性在抗原性上与病毒粒子酶相关;(c) 当蛙病毒在来自广泛不同物种的细胞中生长时,病毒粒子蛋白激酶的抗原和生化特异性保持不变;(d) 对感染不同温度敏感型蛙病毒突变体的细胞进行筛选表明,某些病毒突变体在非允许温度下培养时不能合成这种蛋白激酶。

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