一般酸碱催化对核糖核酸酶A的作用
Value of general Acid-base catalysis to ribonuclease a.
作者信息
Thompson J E, Raines R T
出版信息
J Am Chem Soc. 1994 Jun;116(12):5467-8. doi: 10.1021/ja00091a060.
Abstract
Bovine pancreatic ribonuclease A (RNase A) is a much studied enzyme that efficiently catalyzes the cleavage of RNA. The active site of RNase A contains two histidine residues with imidazole groups positioned to act as a general base (H12) and a general acid (H119) during catalysis of RNA cleavage. Recombinant DNA techniques were used to produce mutant enzymes in which either H12 or H119 was replaced with an alanine residue. Each mutation resulted in a 10-fold decrease in the value of / for cleaving either poly(C) or UpA. Thus, H12 and H119 each lower by 5–6 kcal/mol the free energy of the rate-limiting transition state during RNA cleavage. The value of / for cleavage of UpOCH--NO was decreased by 10-fold by replacing H12 but was unaffected by replacing H119. This result provides the first direct evidence that H119 acts as a general acid during catalysis by RNase A.
摘要
牛胰核糖核酸酶A（RNase A）是一种经过大量研究的酶，它能高效催化RNA的切割。RNase A的活性位点包含两个组氨酸残基，其咪唑基团在RNA切割催化过程中分别作为广义碱（H12）和广义酸（H119）发挥作用。利用重组DNA技术制备了突变酶，其中H12或H119被丙氨酸残基取代。每种突变导致切割聚（C）或UpA的kcat/Km值降低10倍。因此，H12和H119在RNA切割过程中各自将限速过渡态的自由能降低了5 - 6千卡/摩尔。通过替换H12，UpOCH₂NO的切割kcat/Km值降低了10倍，但替换H119对其没有影响。这一结果首次直接证明了H119在RNase A催化过程中作为广义酸发挥作用。
相似文献
1
Value of general Acid-base catalysis to ribonuclease a.一般酸碱催化对核糖核酸酶A的作用
J Am Chem Soc. 1994 Jun;116(12):5467-8. doi: 10.1021/ja00091a060.
2
Histidine-40 of ribonuclease T1 acts as base catalyst when the true catalytic base, glutamic acid-58, is replaced by alanine.当核糖核酸酶T1的真正催化碱基谷氨酸-58被丙氨酸取代时，其组氨酸-40充当碱基催化剂。
Biochemistry. 1990 Sep 25;29(38):9064-72. doi: 10.1021/bi00490a025.
3
Energetics of ribonuclease A catalysis. 1. pH, ionic strength, and solvent isotope dependence of the hydrolysis of cytidine cyclic 2',3'-phosphate.核糖核酸酶A催化的能量学。1. 胞苷2',3'-环磷酸水解的pH值、离子强度和溶剂同位素依赖性。
Biochemistry. 1983 Oct 25;22(22):5123-34. doi: 10.1021/bi00291a011.
4
Three-dimensional structure of ribonuclease Ms*3'-guanylic acid complex at 2.5 A resolution.核糖核酸酶Ms与3'-鸟苷酸复合物在2.5埃分辨率下的三维结构。
FEBS Lett. 1991 Jun 3;283(2):207-9. doi: 10.1016/0014-5793(91)80589-u.
5
Kinetic Isotope Effects and Stereochemical Studies on a Ribonuclease Model: Hydrolysis Reactions of Uridine 3'-Nitrophenyl Phosphate.核糖核酸酶模型的动力学同位素效应和立体化学研究：尿苷3'-硝基苯磷酸酯的水解反应
Bioorg Chem. 2000 Jun;28(3):119-133. doi: 10.1006/bioo.2000.1170.
6
His...Asp catalytic dyad of ribonuclease A: conformational stability of the wild-type, D121N, D121A, and H119A enzymes.他的……核糖核酸酶A的天冬氨酸催化二元体：野生型、D121N、D121A和H119A酶的构象稳定性。
Biochemistry. 1998 Dec 22;37(51):17958-64. doi: 10.1021/bi981688j.
7
Role of aspartic acid 121 in human pancreatic ribonuclease catalysis.天冬氨酸121在人胰腺核糖核酸酶催化中的作用。
Mol Cell Biochem. 2005 Jul;275(1-2):95-101. doi: 10.1007/s11010-005-0997-8.
8
Integration of kinetic isotope effect analyses to elucidate ribonuclease mechanism.整合动力学同位素效应分析以阐明核糖核酸酶机制。
Biochim Biophys Acta. 2015 Nov;1854(11):1801-8. doi: 10.1016/j.bbapap.2015.04.022. Epub 2015 Apr 30.
9
Intramolecular acid-base catalysis of a phosphate diester: modeling the ribonuclease mechanism.磷酸二酯的分子内酸碱催化：核糖核酸酶机制的建模
J Am Chem Soc. 2008 Feb 27;130(8):2436-7. doi: 10.1021/ja710693x. Epub 2008 Feb 5.
10
Decavanadate inhibits catalysis by ribonuclease A.十钒酸盐抑制核糖核酸酶A的催化作用。
Arch Biochem Biophys. 2000 Sep 1;381(1):25-30. doi: 10.1006/abbi.2000.1951.
引用本文的文献
1
Glycosylation of Structured Protein Domains in Cell-Free Reaction Environments.无细胞反应环境中结构化蛋白质结构域的糖基化
ACS Synth Biol. 2025 Jun 20;14(6):2354-2367. doi: 10.1021/acssynbio.5c00229. Epub 2025 May 28.
2
Mechanism-Based Inactivation of Ribonuclease A.基于机制的核糖核酸酶A失活
J Org Chem. 1995 Oct;60(21):6930-6936. doi: 10.1021/jo00126a051.
3
Mechanism of activation of contact-dependent growth inhibition tRNase toxin by the amino acid biogenesis factor CysK in the bacterial competition system.细菌竞争系统中氨基酸生物合成因子CysK激活接触依赖性生长抑制tRNase毒素的机制。
Nucleic Acids Res. 2025 Jan 7;53(1). doi: 10.1093/nar/gkae735.
4
Accurately Predicting Protein p Values Using Nonequilibrium Alchemy.使用非平衡炼金术准确预测蛋白质 p 值。
J Chem Theory Comput. 2023 Nov 14;19(21):7833-7845. doi: 10.1021/acs.jctc.3c00721. Epub 2023 Oct 11.
5
Semisynthesis of Human Ribonuclease-S.人核糖核酸酶-S 的半合成。
Bioconjug Chem. 2021 Jan 20;32(1):82-87. doi: 10.1021/acs.bioconjchem.0c00557. Epub 2020 Dec 9.
6
Antimicrobial Synergy of a Ribonuclease and a Peptide Secreted by Human Cells.一种核糖核酸酶与人类细胞分泌的一种肽的抗菌协同作用。
ACS Infect Dis. 2020 Nov 13;6(11):3083-3088. doi: 10.1021/acsinfecdis.0c00594. Epub 2020 Oct 15.
7
RNA promotes phase separation of glycolysis enzymes into yeast G bodies in hypoxia.RNA 促进糖酵解酶在低氧条件下形成酵母 G 体的相分离。
Elife. 2020 Apr 16;9:e48480. doi: 10.7554/eLife.48480.
8
A catalytic protein-proteomimetic complex: using aromatic oligoamide foldamers as activators of RNase S.一种催化性蛋白质模拟复合物：使用芳香族寡酰胺折叠体作为核糖核酸酶S的激活剂。
Chem Sci. 2019 Feb 21;10(14):3956-3962. doi: 10.1039/c9sc00374f. eCollection 2019 Apr 14.
9
Generation of 2',3'-Cyclic Phosphate-Containing RNAs as a Hidden Layer of the Transcriptome.生成含2',3'-环磷酸酯的RNA作为转录组的隐藏层。
Front Genet. 2018 Nov 27;9:562. doi: 10.3389/fgene.2018.00562. eCollection 2018.
10
Knockout of the Ribonuclease Inhibitor Gene Leaves Human Cells Vulnerable to Secretory Ribonucleases.核糖核酸酶抑制剂基因敲除使人类细胞易受分泌型核糖核酸酶的影响。
Biochemistry. 2016 Nov 22;55(46):6359-6362. doi: 10.1021/acs.biochem.6b01003. Epub 2016 Nov 8.
本文引用的文献
1
The active site and mechanism of action of bovine pancreatic ribonuclease. 5. The charge types at the active centre.牛胰核糖核酸酶的活性位点与作用机制。5. 活性中心的电荷类型。
Biochem J. 1962 Oct;85(1):139-44. doi: 10.1042/bj0850139.
2
On the aggregation of bovine pancreatic ribonuclease.关于牛胰核糖核酸酶的聚集
Arch Biochem Biophys. 1962 Sep;Suppl 1:217-22.
3
The active site and mechanism of action of bovine pancreatic ribonuclease. 3. The pH-dependence of the kinetic parameters for the hydrolysis of cytidine 2',3'-phosphate.牛胰核糖核酸酶的活性位点及作用机制。3. 2',3'-磷酸胞苷水解动力学参数的pH依赖性。
Biochem J. 1962 Oct;85(1):127-34. doi: 10.1042/bj0850127.
4
A spectrophotometric assay for ribonuclease activity using cytidylyl-(3',5')-adenosine and uridylyl-(3',5')-adenosine as substrates.一种以胞苷酰-(3',5')-腺苷和尿苷酰-(3',5')-腺苷为底物的核糖核酸酶活性分光光度测定法。
FEBS Lett. 1968 Jul;1(1):29-31. doi: 10.1016/0014-5793(68)80010-9.
5
Understanding the rates of certain enzyme-catalyzed reactions: proton abstraction from carbon acids, acyl-transfer reactions, and displacement reactions of phosphodiesters.了解某些酶催化反应的速率：从碳酸中夺取质子、酰基转移反应以及磷酸二酯的取代反应。
Biochemistry. 1993 Nov 16;32(45):11943-52. doi: 10.1021/bi00096a001.
6
Structural determinants of enzymatic processivity.酶促持续合成能力的结构决定因素。
Biochemistry. 1994 May 24;33(20):6031-7. doi: 10.1021/bi00186a001.
7
Energetics of catalysis by ribonucleases: fate of the 2',3'-cyclic phosphodiester intermediate.核糖核酸酶催化的能量学：2',3'-环磷酸二酯中间体的命运
Biochemistry. 1994 Jun 14;33(23):7408-14. doi: 10.1021/bi00189a047.
8
Energetics of ribonuclease A catalysis. 1. pH, ionic strength, and solvent isotope dependence of the hydrolysis of cytidine cyclic 2',3'-phosphate.核糖核酸酶A催化的能量学。1. 胞苷2',3'-环磷酸水解的pH值、离子强度和溶剂同位素依赖性。
Biochemistry. 1983 Oct 25;22(22):5123-34. doi: 10.1021/bi00291a011.
9
Tertiary structure of ribonuclease.核糖核酸酶的三级结构。
Nature. 1967 Mar 4;213(5079):862-5. doi: 10.1038/213862a0.
10
Structure of ribonuclease.核糖核酸酶的结构。
Nature. 1967 Feb 11;213(5076):557-62. doi: 10.1038/213557a0.
Zotero 插件