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聚乙二醇在低酶浓度下增强核糖核苷酸还原酶活性。

Enhancement of ribonucleotide reductase activity at low enzyme concentrations by polyethylene glycol.

作者信息

Kucera R, Paulus H

机构信息

Department of Metabolic Regulation, Boston Biomedical Research Institute, Boston, Mass.

出版信息

Enzyme. 1988;39(2):115-8.

PMID:2456207
Abstract

The estimation of ribonucleotide reductase in cell extracts has been problematical on account of abnormally low activities at low enzyme concentrations, presumably due to subunit dissociation. This problem can be alleviated by assaying the enzyme in the presence of polyethylene glycol. The presence of 15% polyethylene glycol during the assay greatly stimulated ribonucleotide reductase activity at low enzyme concentrations and allowed measurement of enzyme activity in as little as 10(5) mouse L929 cells, a 30-fold enhancement of assay sensitivity. Enzyme activity measured in the presence of 15% polyethylene glycol was proportional to enzyme concentration, thus making possible the accurate measurement of very low levels of ribonucleotide reductase.

摘要

由于在低酶浓度下活性异常低,推测是由于亚基解离,因此在细胞提取物中估算核糖核苷酸还原酶一直存在问题。通过在聚乙二醇存在下测定该酶,可以缓解这个问题。在测定过程中加入15%的聚乙二醇,在低酶浓度下极大地刺激了核糖核苷酸还原酶的活性,使得在仅10⁵个小鼠L929细胞中就能测量酶活性,测定灵敏度提高了30倍。在15%聚乙二醇存在下测得的酶活性与酶浓度成正比,从而使得准确测量极低水平的核糖核苷酸还原酶成为可能。

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