大鼠和非洲爪蟾肝脏中γ-谷氨酰半胱氨酸合成酶的测定、纯化、性质及作用机制
Assay, purification, properties and mechanism of action of gamma-glutamylcysteine synthetase from the liver of the rat and Xenopus laevis.
作者信息
Davis J S, Balinsky J B, Harington J S, Shepherd J B
出版信息
Biochem J. 1973 Aug;133(4):667-78. doi: 10.1042/bj1330667.
Abstract
- An improved radioassay for glutathione synthetase and gamma-glutamylcysteine synthetase was developed. 2. Xenopus laevis liver gamma-glutamylcysteine synthetase was purified 324-fold by saline-bicarbonate extraction, protamine sulphate precipitation, CM-cellulose and DEAE-cellulose column chromatography, and gel filtration. 3. Rat liver gamma-glutamylcysteine synthetase was purified 11400-fold by a procedure similar to that employed for the Xenopus laevis enzyme. 4. Rat liver gamma-glutamylcysteine synthetase activity was inhibited by GSH and activated by glycine. These effects, which were not found in the enzyme from Xenopus laevis, may have a regulatory significance. 5. Isotope-exchange experiments revealed fundamental differences in the partial reactions catalysed by the rat and Xenopus laevis synthetases. The enzyme from Xenopus laevis appears to follow a Bi Bi Uni Uni Ping Pong mechanism, with glutamyl-enzyme as intermediate before the addition of cysteine and the release of gamma-glutamylcysteine. The results for the rat liver enzyme are consistent with a Tri Tri sequential mechanism.
摘要
- 开发了一种改进的谷胱甘肽合成酶和γ-谷氨酰半胱氨酸合成酶放射分析方法。2. 非洲爪蟾肝脏γ-谷氨酰半胱氨酸合成酶通过碳酸氢盐盐水提取、硫酸鱼精蛋白沉淀、CM-纤维素和DEAE-纤维素柱色谱以及凝胶过滤进行纯化，纯化了324倍。3. 大鼠肝脏γ-谷氨酰半胱氨酸合成酶通过与用于非洲爪蟾酶的类似方法纯化了11400倍。4. 大鼠肝脏γ-谷氨酰半胱氨酸合成酶活性受到谷胱甘肽抑制，并被甘氨酸激活。这些在非洲爪蟾酶中未发现的效应可能具有调节意义。5. 同位素交换实验揭示了大鼠和非洲爪蟾合成酶催化的部分反应的根本差异。非洲爪蟾的酶似乎遵循双底物双产物单底物单产物乒乓机制，在添加半胱氨酸和释放γ-谷氨酰半胱氨酸之前，以谷氨酰酶作为中间体。大鼠肝脏酶的结果与三三顺序机制一致。
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本文引用的文献
1
Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
J Biol Chem. 1951 Nov;193(1):265-75.
2
Enzymatic synthesis of glutathione.谷胱甘肽的酶促合成。
J Biol Chem. 1951 Jan;188(1):221-40.
3
Enzymatic synthesis of gamma-glutamylcysteine.γ-谷氨酰半胱氨酸的酶促合成
J Biol Chem. 1955 Jun;214(2):639-46.
4
The kinetics of enzyme-catalyzed reactions with two or more substrates or products. I. Nomenclature and rate equations.具有两种或更多种底物或产物的酶催化反应动力学。I. 命名法和速率方程。
Biochim Biophys Acta. 1963 Jan 8;67:104-37. doi: 10.1016/0006-3002(63)91800-6.
5
Acidic peptides of the lens. 6. Metabolism of gamma-glutamyl peptides in subcellular fractions of rabbit liver.晶状体的酸性肽。6. 兔肝脏亚细胞组分中γ-谷氨酰肽的代谢
Biochem J. 1961 Apr;79(1):118-28. doi: 10.1042/bj0790118.
6
Peptidebond synthesis in higher plants. II. Studies on the mechanism of synthesis of gamma-glutamylcysteine.高等植物中的肽键合成。II. γ-谷氨酰半胱氨酸合成机制的研究。
Arch Biochem Biophys. 1954 Sep;52(1):22-32. doi: 10.1016/0003-9861(54)90085-5.
7
Peptide-bond synthesis in higher plants. I. The synthesis of glutathione.高等植物中的肽键合成。I. 谷胱甘肽的合成。
Arch Biochem Biophys. 1953 Dec;47(2):241-50. doi: 10.1016/0003-9861(53)90464-0.
8
Synthesis of glutathione from gamma-glutamylcysteine.由γ-谷氨酰半胱氨酸合成谷胱甘肽。
J Biol Chem. 1953 Apr;201(2):573-86.
9
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J Biol Chem. 1953 Apr;201(2):561-71.
10
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