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乳糖阻遏蛋白与聚d(A-T)的结合：复合物的OD和CD熔解

Binding of lactose repressor to poly d(A-T) : OD AND CD melting of the complex.

作者信息

Clement R, Daune M P

出版信息

Nucleic Acids Res. 1975 Mar;2(3):303-18. doi: 10.1093/nar/2.3.303.

DOI:10.1093/nar/2.3.303
PMID:1093136
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC342837/
Abstract

The binding of lactose repressor to poly d(A-T) at low ionic strength has been investigated by heat denaturation. The poly d(A-T) melting is monitored by optical density and the protein melting by circular dichroism. From the modification of the poly d(A-T) melting curve we estimate a maximum binding ratio of about one tetrameric repressor to about 20 basic pairs. The repressor melting can be interpreted as a global shift from a to b structure of about 25 residues per subunit. The melting curves of poly d(A-T) and repressor have not a shape easy to interpret; nevertheless both show a cooperative transition in the same temperature range where we can evaluate that about 3.8 aminoacid residues shift from a to b structure when 1 bases pair melt.

摘要

通过热变性研究了低离子强度下乳糖阻遏物与聚d(A-T)的结合。通过光密度监测聚d(A-T)的解链，通过圆二色性监测蛋白质的解链。根据聚d(A-T)解链曲线的变化，我们估计最大结合比约为一个四聚体阻遏物与约20个碱基对结合。阻遏物的解链可解释为每个亚基约25个残基从a结构到b结构的整体转变。聚d(A-T)和阻遏物的解链曲线形状不易解释；然而，两者在相同温度范围内均显示出协同转变，在此温度范围内我们可以评估出当1个碱基对解链时约3.8个氨基酸残基从a结构转变为b结构。

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Nucleic Acids Res. 1981 Dec 11;9(23):6505-25. doi: 10.1093/nar/9.23.6505.
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Lac repressor purification without inactivation of DNA binding activity.不使DNA结合活性失活的乳糖阻遏物纯化。
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本文引用的文献

1
Effect of reconstitution conditions on the structure of Escherichia coli 30-S ribosomol-subunit components.重构条件对大肠杆菌30-S核糖体亚基组分结构的影响。
Eur J Biochem. 1974 Nov 1;49(1):185-94. doi: 10.1111/j.1432-1033.1974.tb03824.x.
2
The lac repressor protein: molecular shape, subunit structure, and proposed model for operator interaction based on structural studies of microcrystals.乳糖阻遏蛋白：基于微晶结构研究的分子形状、亚基结构及与操纵基因相互作用的模型
Proc Natl Acad Sci U S A. 1974 Mar;71(3):593-7. doi: 10.1073/pnas.71.3.593.
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The nucleotide sequence of the lac operator.乳糖操纵基因的核苷酸序列。
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3581-4. doi: 10.1073/pnas.70.12.3581.
4
The amino-acid sequence of lac repressor.乳糖阻遏蛋白的氨基酸序列。
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3576-80. doi: 10.1073/pnas.70.12.3576.
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Magnetic circular dichroism studies. XIX. Determination of the tyrosine: tryptophan ratio in proteins.磁圆二色性研究。第十九部分。蛋白质中酪氨酸与色氨酸比例的测定。
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Equilibrium and kinetic studies of Escherichia coli lac repressor-inducer interactions.大肠杆菌乳糖阻遏物与诱导剂相互作用的平衡和动力学研究。
J Mol Biol. 1972 Apr 28;66(1):143-55. doi: 10.1016/s0022-2836(72)80012-3.
7
Secondary structure of the lac repressor.乳糖阻遏蛋白的二级结构。
Biochem Biophys Res Commun. 1972 Jun 28;47(6):1438-43. doi: 10.1016/0006-291x(72)90233-1.
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Determination of the secondary structures of proteins by circular dichroism and optical rotatory dispersion.通过圆二色性和旋光色散测定蛋白质的二级结构。
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