人红细胞6-磷酸葡萄糖酸脱氢酶的半位点反应活性
Half-site reactivity in 6-phosphogluconate dehydrogenase from human erythrocytes.
作者信息
Dallocchio F, Matteuzzi M, Bellini T
出版信息
Biochem J. 1985 Apr 1;227(1):305-10. doi: 10.1042/bj2270305.
Abstract
6-Phosphogluconate dehydrogenase from human erythrocytes was purified by an improved procedure. Binding studies showed that the dimeric enzyme binds 2 mol of NADP+/mol but only 1 mol of NADPH/mol, and that the bindings of oxidized and reduced coenzyme are mutually exclusive. From initial-rate kinetics and inhibition studies, a sequential random-order mechanism is proposed. Double-reciprocal plots with NADP+ as varied substrate show a downward curvature, indicating a negative co-operativity. We suggest that the negative co-operativity observed kinetically is a result of the half-site reactivity for the NADPH. The different binding stoichiometries for NADP+ and NADPH generate a non-linear relationship between the apparent dissociation constant for the NADPH and the concentrations of the NADP+, resulting in a regulatory mechanism highly sensitive to the changes in the NADP+/NADPH ratio.
摘要
通过改进的方法对人红细胞中的6-磷酸葡萄糖酸脱氢酶进行了纯化。结合研究表明，二聚体酶每摩尔结合2摩尔NADP⁺但每摩尔仅结合1摩尔NADPH，并且氧化型和还原型辅酶的结合是相互排斥的。根据初速率动力学和抑制研究，提出了一种顺序随机顺序机制。以NADP⁺作为可变底物的双倒数图显示向下弯曲，表明存在负协同性。我们认为动力学上观察到的负协同性是NADPH半位点反应性的结果。NADP⁺和NADPH不同的结合化学计量在NADPH的表观解离常数与NADP⁺浓度之间产生非线性关系，从而形成对NADP⁺/NADPH比值变化高度敏感的调节机制。
相似文献
1
Half-site reactivity in 6-phosphogluconate dehydrogenase from human erythrocytes.人红细胞6-磷酸葡萄糖酸脱氢酶的半位点反应活性
Biochem J. 1985 Apr 1;227(1):305-10. doi: 10.1042/bj2270305.
2
[[[Purification and properties of 6-phosphogluconate dehydrogenase from human erythrocytes].[[[人红细胞6-磷酸葡萄糖酸脱氢酶的纯化及性质]。
Boll Soc Ital Biol Sper. 1981 Oct 30;57(20):2018-21.
3
Human brain 6-phosphogluconate dehydrogenase: purification and kinetic properties.
J Neurochem. 1985 Feb;44(2):510-7. doi: 10.1111/j.1471-4159.1985.tb05443.x.
4
Effect of the substrate on the binding of coenzyme and coenzyme analogues to 6-phosphogluconate dehydrogenase from Candida utilis.底物对辅酶及辅酶类似物与产朊假丝酵母6-磷酸葡萄糖酸脱氢酶结合的影响。
J Biol Chem. 1981 Nov 10;256(21):10778-80.
5
Kinetic studies of 6-phosphogluconate dehydrogenase from sheep liver.绵羊肝脏6-磷酸葡萄糖酸脱氢酶的动力学研究
Eur J Biochem. 1986 May 2;156(3):555-67. doi: 10.1111/j.1432-1033.1986.tb09615.x.
6
6-Phosphogluconate dehydrogenase: the mechanism of action investigated by a comparison of the enzyme from different species.6-磷酸葡萄糖酸脱氢酶：通过比较不同物种的酶来研究其作用机制。
Biochim Biophys Acta. 1998 Dec 8;1429(1):83-92. doi: 10.1016/s0167-4838(98)00222-2.
7
Crystallographic study of coenzyme, coenzyme analogue and substrate binding in 6-phosphogluconate dehydrogenase: implications for NADP specificity and the enzyme mechanism.6-磷酸葡萄糖酸脱氢酶中辅酶、辅酶类似物及底物结合的晶体学研究：对NADP特异性及酶作用机制的启示
Structure. 1994 Jul 15;2(7):651-68. doi: 10.1016/s0969-2126(00)00066-6.
8
Human granulocyte 6 phosphogluconate dehydrogenase. Purification by elective elution with NADP+, immunological and kinetic properties.人粒细胞6-磷酸葡萄糖酸脱氢酶。用NADP+选择性洗脱进行纯化、免疫学及动力学性质
Biochimie. 1975;57(3):325-35. doi: 10.1016/s0300-9084(75)80308-7.
9
Thermodynamic characterization of substrate and inhibitor binding to Trypanosoma brucei 6-phosphogluconate dehydrogenase.底物和抑制剂与布氏锥虫6-磷酸葡萄糖酸脱氢酶结合的热力学特征
FEBS J. 2007 Dec;274(24):6426-35. doi: 10.1111/j.1742-4658.2007.06160.x. Epub 2007 Nov 15.
10
[Kinetic characteristics of 6-phosphogluconate dehydrogenase from bull adrenal cortex].
Ukr Biokhim Zh (1978). 1989 Sep-Oct;61(5):92-5.
引用本文的文献
1
6-Phosphogluconate dehydrogenase and its crystal structures.6-磷酸葡萄糖酸脱氢酶及其晶体结构。
Acta Crystallogr F Struct Biol Commun. 2022 Mar 1;78(Pt 3):96-112. doi: 10.1107/S2053230X22001091. Epub 2022 Feb 23.
2
Kinetic and spectroscopic evidence of negative cooperativity in the action of lysine 2,3-aminomutase.赖氨酸 2,3-氨基变位酶作用中的负协同性的动力学和光谱证据。
J Phys Chem B. 2010 Dec 16;114(49):16118-24. doi: 10.1021/jp103856m. Epub 2010 Jul 7.
3
Kinetic and X-ray structural evidence for negative cooperativity in substrate binding to nicotinate mononucleotide adenylyltransferase (NMAT) from Bacillus anthracis.炭疽芽孢杆菌烟酸单核苷酸腺苷酰转移酶（NMAT）底物结合中负协同性的动力学和X射线结构证据。
J Mol Biol. 2009 Jan 23;385(3):867-88. doi: 10.1016/j.jmb.2008.10.037. Epub 2008 Oct 19.
4
Kinetic properties of hexose-monophosphate dehydrogenases. II. Isolation and partial purification of 6-phosphogluconate dehydrogenase from rat liver and kidney cortex.己糖单磷酸脱氢酶的动力学特性。II. 大鼠肝脏和肾皮质中6-磷酸葡萄糖酸脱氢酶的分离及部分纯化
Mol Cell Biochem. 1995 Mar 23;144(2):97-104. doi: 10.1007/BF00944387.
本文引用的文献
1
[Kinetic properties of D-gluconate-6-phosphate dehydrogenase and the activating effect of the magnesium ion].[6-磷酸-D-葡萄糖酸脱氢酶的动力学特性及镁离子的激活作用]
Arch Sci Biol (Bologna). 1962 Jan-Mar;46:83-91.
2
Estimation of dissociation constant of enzyme-ligand complex from fluorometric data by "difference" method.用“差值”法根据荧光数据估算酶-配体复合物的解离常数。
FEBS Lett. 1972 Jan 1;19(4):308-310. doi: 10.1016/0014-5793(72)80067-x.
3
Antagonistic homotropic interactions as a possible explanation of coenzyme activation of glutamate dehydrogenase.拮抗同促相互作用作为谷氨酸脱氢酶辅酶激活的一种可能解释。
FEBS Lett. 1968 Oct;1(5):349-352. doi: 10.1016/0014-5793(68)80153-x.
4
The molecular weight and subunit structure of human erythrocyte 6-phosphogluconate dehydrogenase.人红细胞6-磷酸葡萄糖酸脱氢酶的分子量及亚基结构。
Eur J Biochem. 1974 Feb 15;42(1):225-35. doi: 10.1111/j.1432-1033.1974.tb03332.x.
5
Crystalline 6-phosphogluconate dehydrogenase from sheep liver.来自绵羊肝脏的结晶6-磷酸葡萄糖酸脱氢酶。
Eur J Biochem. 1973 Oct 5;38(2):229-38. doi: 10.1111/j.1432-1033.1973.tb03054.x.
6
Sheep liver 6-phosphogluconate dehydrogenase: isolation procedure and effect of pH, ionic strength, and metal ions on the kinetic parameters.绵羊肝脏6-磷酸葡萄糖酸脱氢酶：分离方法以及pH、离子强度和金属离子对动力学参数的影响
Arch Biochem Biophys. 1973 Feb;154(2):623-35. doi: 10.1016/0003-9861(73)90017-9.
7
Human erythrocyte glucose 6-phosphate dehydrogenase. Interaction with oxidized and reduced coenzyme.人类红细胞葡萄糖6-磷酸脱氢酶。与氧化型和还原型辅酶的相互作用。
Biochem Biophys Res Commun. 1974 Oct 8;60(3):999-1005. doi: 10.1016/0006-291x(74)90412-4.
8
Human erythrocyte glucose 6-phosphate dehydrogenase. Content of bound coenzyme.人红细胞葡萄糖6-磷酸脱氢酶。结合辅酶含量。
Biochem Biophys Res Commun. 1974 Jul 10;59(1):406-13. doi: 10.1016/s0006-291x(74)80221-4.
9
Human 6-phosphogluconate dehydrogenase. Purification of the erythrocyte enzyme and the influence of ions and NADPH on its activity.人6-磷酸葡萄糖酸脱氢酶。红细胞酶的纯化以及离子和NADPH对其活性的影响。
Eur J Biochem. 1974 Feb 15;42(1):213-23. doi: 10.1111/j.1432-1033.1974.tb03331.x.
10
Inhibition of co-operative enzymes by substrate-analogues: possible implications for the physiological significance of negative co-operativity illustrated by phenylalanine metabolism in higher plants.底物类似物对协同酶的抑制作用：以高等植物苯丙氨酸代谢为例探讨负协同性生理意义的可能影响。
J Theor Biol. 1976 Aug 7;60(2):441-7. doi: 10.1016/0022-5193(76)90070-9.
Zotero 插件