负同促协同性与亲和力异质性：制备具有最大亲和力同质性的酵母甘油醛-3-磷酸脱氢酶。

Negative homotropic cooperativity and affinity heterogeneity: preparation of yeast glyceraldehyde-3-phosphate dehydrogenase with maximal affinity homogeneity.

作者信息

Gennis L S

出版信息

Proc Natl Acad Sci U S A. 1976 Nov;73(11):3928-32. doi: 10.1073/pnas.73.11.3928.

DOI:10.1073/pnas.73.11.3928

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC431269/

Abstract

A three-step procedure including affinity chromatography on NAD+-azobenzamidopropyl-Sepharose has been designed for the purification of yeast glyceraldehyde-3-phosphate dehydrogenase [D-glyceraldehyde-3-phosphate: NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12] with maximized specific activity and maximized homogeneity with respect to affinity for the coenzyme, NAD+. Binding isotherms allow the analysis of cooperativity patterns that disclose both the average ligand affinity in the system and the distribution of ligands among the sites, only for systems with complete affinity homogeneity. The presence of affinity heterogeneity, resulting from multiple oligomeric species differing only in their affinity for coenzyme, gives rise to isotherms which falsely manifest apparent negative cooperativity. A method for distinguishing negative homotropic cooperativity from affinity heterogeneity is suggested.

摘要

已设计出一种三步法程序，其中包括在NAD⁺-偶氮苯甲酰胺丙基-琼脂糖凝胶上进行亲和色谱，用于纯化酵母甘油醛-3-磷酸脱氢酶[D-甘油醛-3-磷酸：NAD⁺氧化还原酶（磷酸化），EC 1.2.1.12]，使其比活性最大化，并且在对辅酶NAD⁺的亲和力方面具有最大的均一性。结合等温线允许分析协同模式，这些模式揭示了系统中的平均配体亲和力以及配体在位点之间的分布情况，这仅适用于具有完全亲和力均一性的系统。由仅对辅酶亲和力不同的多种寡聚体物种导致的亲和力异质性的存在，会产生错误显示出明显负协同性的等温线。本文提出了一种区分负同促协同性与亲和力异质性的方法。

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本文引用的文献

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Proc Natl Acad Sci U S A. 1955 Jun 15;41(6):327-38. doi: 10.1073/pnas.41.6.327.

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THE COMPARATIVE ENZYMOLOGY OF LACTIC DEHYDROGENASES. I. PROPERTIES OF THE CRYSTALLINE BEEF AND CHICKEN ENZYMES.乳酸脱氢酶的比较酶学。I. 结晶牛肉和鸡肉酶的性质。

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