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首页> 外文期刊>Journal of Virology >Biochemical characterization of equine herpesvirus type 3-induced deoxythymidine kinase purified from lytically infected horse embryo dermal fibroblasts.
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Biochemical characterization of equine herpesvirus type 3-induced deoxythymidine kinase purified from lytically infected horse embryo dermal fibroblasts.

机译：纯化马胚胎皮毛细胞纯化的马疱疹病毒3型诱导的脱氧肾上腺素激酶的生化特征。

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摘要

Infection of horse KyED cells with equine herpesvirus type 3 (EHV-3) resulted in a sevenfold increase in cytosol deoxythymidine kinase (dTK) activity. The EHV-3 dTK was purified from KyED cytosol dTK by affinity chromatography on deoxythymidine-Sepharose and characterized with respect to its electrophoretic mobility, molecular weight, substrate specificity, phosphate donor specificity, and immunological specificity. The purified EHV-3 dTK migrated in polyacrylamide gels with an Rf of 0.30 and sedimented in glycerol gradients with an S value of 5.13, corresponding to a molecular weight of 83,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielded a single band with a molecular weight of 38,000 to 40,000. Antiserum prepared against the EHV-3 dTK induced in KyED cells neutralized the EHV-3-induced enzyme activity but not the dTK purified from uninfected cells. EHV-3 dTK was less sensitive to feedback inhibition to dTTP and had a lower Ki for the antiviral compound 1-beta-D-arabinofuranyosylthymine and a lower Km for the substrate deoxythymidine. These results indicate that infection of cells with EHV-3 results in the induction of a new virus-coded dTK activity which meets the criteria of Jensen for an evolutionary primitive enzyme.
机译：用马卵黄素3（EHV-3）感染马ky细胞，导致细胞溶溶氧脱氧激酶（DTK）活性七倍增加。通过亲和层析在脱氧血清 - 琼脂糖上的亲和色谱法从Kyed Cytosol DTK纯化EHV-3 DTK，并相对于其电泳迁移率，分子量，底物特异性，磷酸盐给药特异性和免疫特异性。纯化的EHV-3DTK在聚丙烯酰胺凝胶中迁移，RF为0.30，并在甘油梯度中沉积，S值为5.13，对应于83,000的分子量。十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳产生单个带，分子量为38,000至40,000。抗血清对Kyed细胞诱导的EHV-3 DTK中和中和EHV-3诱导的酶活性，但不是从未感染的细胞纯化的DTK。 EHV-3 DTK对DTTP的反馈抑制不太敏感，并且对于抗病毒化合物1-Beta-d-Arabinofurany ranyosylynine和基材脱氧尿苷的低km具有较低的Ki。这些结果表明，具有EHV-3的细胞的感染导致诱导符合Jensen的标准的新病毒编码的DTK活性。

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