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从肺炎球菌中提取的脱氧核糖核酸-膜组分在聚丙烯酰胺凝胶上的酶原位活性。

In situ activity of enzymes on polyacrylamide gels of a deoxyribonucleic acid-membrane fraction extracted from pneumococci.

作者信息

Firshein W

出版信息

J Bacteriol. 1974 Jun;118(3):1101-10. doi: 10.1128/jb.118.3.1101-1110.1974.

DOI:10.1128/jb.118.3.1101-1110.1974
PMID:4151452
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC246861/
Abstract

A deoxyribonucleic acid (DNA)-membrane fraction extracted from Diplococcus pneumoniae was subjected to polyacrylamide gel electrophoresis after treatment with 0.16% sodium dodecyl sulfate. At least two DNA polymerase activities were detected by in situ assays with appropriate substrates, templates, and inhibitors, including a co-polymer of deoxyadenylic and thymidylic acid and N-ethylmaleimide. This activity coincided with a fraction in the gel containing 7.5, 9.4, and 24%, respectively of the DNA, phospholipid, and protein present in the DNA-membrane fraction before electrophoresis and sodium dodecyl sulfate treatment. Assays with minced gels showed that several nuclease activities, deoxyribonucleotide kinase activity, and DNA ligase activity also coincided with this fraction. However, ribonucleoside diphosphate reductase activity did not. These results demonstrate that a complex of enzymes involved in DNA replication is firmly bound to the DNA-membrane fraction in pneumococci.

摘要

从肺炎双球菌中提取的脱氧核糖核酸（DNA）-膜组分，在用0.16%十二烷基硫酸钠处理后进行聚丙烯酰胺凝胶电泳。通过使用合适的底物、模板和抑制剂（包括脱氧腺苷酸和胸苷酸的共聚物以及N-乙基马来酰亚胺）进行原位测定，检测到至少两种DNA聚合酶活性。这种活性与凝胶中的一个组分相对应，该组分在电泳和十二烷基硫酸钠处理之前的DNA-膜组分中分别含有7.5%、9.4%和24%的DNA、磷脂和蛋白质。对切碎凝胶的测定表明，几种核酸酶活性、脱氧核糖核苷酸激酶活性和DNA连接酶活性也与该组分相对应。然而，核糖核苷二磷酸还原酶活性并不对应。这些结果表明，参与DNA复制的酶复合物牢固地结合在肺炎球菌的DNA-膜组分上。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a58f/246861/3c8d7c92a7d5/jbacter00342-0350-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a58f/246861/3c8d7c92a7d5/jbacter00342-0350-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a58f/246861/3c8d7c92a7d5/jbacter00342-0350-a.jpg

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本文引用的文献

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Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
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Tetracycline inhibits propagation of deoxyribonucleic acid replication and alters membrane properties.四环素抑制脱氧核糖核酸复制的传播并改变膜的性质。
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Alterations of deoxyribonucleoside triphosphate pools in Escherichia coli: effects on deoxyribonucleic acid replication and evidence for compartmentation.大肠杆菌中三磷酸脱氧核糖核苷库的改变：对脱氧核糖核酸复制的影响及区室化证据。
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Glycolipids stimulate DNA polymerase activity in a DNA-membrane fraction and in a partially purified polymerase system extracted from pneumococci.糖脂可刺激肺炎球菌提取的DNA-膜组分和部分纯化的聚合酶系统中的DNA聚合酶活性。
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