单点突变（F429H）将酶 CYP2B4 转化为血红素加氧酶：QM/MM 研究。
A single-site mutation (F429H) converts the enzyme CYP 2B4 into a heme oxygenase: a QM/MM study.
机构信息
Institute of Chemistry and the Lise Meitner-Minerva Center for Computational Quantum Chemistry, The Hebrew University of Jerusalem, 91904 Jerusalem, Israel.
出版信息
J Am Chem Soc. 2012 Mar 7;134(9):4053-6. doi: 10.1021/ja211905e. Epub 2012 Feb 22.
Abstract
The intriguing deactivation of the cytochrome P450 (CYP) 2B4 enzyme induced by mutation of a single residue, Phe429 to His, is explored by quantum mechanical/molecular mechanical calculations of the O-OH bond activation of the (Fe(3+)OOH)(-) intermediate. It is found that the F429H mutant of CYP 2B4 undergoes homolytic instead of heterolytic O-OH bond cleavage. Thus, the mutant acquires the following characteristics of a heme oxygenase enzyme: (a) donation by His429 of an additional NH---S H-bond to the cysteine ligand combined with the presence of the substrate retards the heterolytic cleavage and gives rise to homolytic O-OH cleavage, and (b) the Thr302/water cluster orients nascent OH(•) and ensures efficient meso hydroxylation.
摘要
通过对（Fe(3+)OOH)(-）中间物的 O-OH 键活化的量子力学/分子力学计算，探究了单个残基 Phe429 突变为 His 后细胞色素 P450（CYP）2B4 酶的有趣失活。结果发现，CYP 2B4 的 F429H 突变体经历均裂而不是异裂 O-OH 键裂解。因此，突变体获得了血红素加氧酶酶的以下特征：(a) His429 向半胱氨酸配体提供额外的 NH---S H 键结合与底物的存在，延缓了异裂裂解并导致均裂 O-OH 裂解，和 (b) Thr302/水簇使新生的 OH(•)取向并确保有效的间位羟化。
相似文献
1
A single-site mutation (F429H) converts the enzyme CYP 2B4 into a heme oxygenase: a QM/MM study.单点突变（F429H）将酶 CYP2B4 转化为血红素加氧酶：QM/MM 研究。
J Am Chem Soc. 2012 Mar 7;134(9):4053-6. doi: 10.1021/ja211905e. Epub 2012 Feb 22.
2
Structural and functional characterization of a cytochrome P450 2B4 F429H mutant with an axial thiolate-histidine hydrogen bond.具有轴向硫醇盐-组氨酸氢键的细胞色素P450 2B4 F429H突变体的结构与功能表征
Biochemistry. 2014 Aug 12;53(31):5080-91. doi: 10.1021/bi5003794. Epub 2014 Jul 31.
3
Role of the Proximal Cysteine Hydrogen Bonding Interaction in Cytochrome P450 2B4 Studied by Cryoreduction, Electron Paramagnetic Resonance, and Electron-Nuclear Double Resonance Spectroscopy.通过低温还原、电子顺磁共振和电子-核双共振光谱研究近端半胱氨酸氢键相互作用在细胞色素P450 2B4中的作用
Biochemistry. 2016 Feb 16;55(6):869-83. doi: 10.1021/acs.biochem.5b00744. Epub 2016 Feb 3.
4
F429 Regulation of Tunnels in Cytochrome P450 2B4: A Top Down Study of Multiple Molecular Dynamics Simulations.F429对细胞色素P450 2B4中通道的调控：多分子动力学模拟的自上而下研究
PLoS One. 2015 Sep 28;10(9):e0137075. doi: 10.1371/journal.pone.0137075. eCollection 2015.
5
Replacement of active-site cysteine-436 by serine converts cytochrome P450 2B4 into an NADPH oxidase with negligible monooxygenase activity.将活性位点的半胱氨酸-436替换为丝氨酸会将细胞色素P450 2B4转变为一种单加氧酶活性可忽略不计的NADPH氧化酶。
J Inorg Biochem. 2002 Sep 20;91(4):542-53. doi: 10.1016/s0162-0134(02)00438-5.
6
Quantum mechanical/molecular mechanical study of mechanisms of heme degradation by the enzyme heme oxygenase: the strategic function of the water cluster.血红素加氧酶催化血红素降解机制的量子力学/分子力学研究：水簇的关键作用
J Am Chem Soc. 2008 Feb 13;130(6):1953-65. doi: 10.1021/ja076679p. Epub 2008 Jan 18.
7
Abolition of oxygenase function, retention of NADPH oxidase activity, and emergence of peroxidase activity upon replacement of the axial cysteine-436 ligand by histidine in cytochrome P450 2B4.细胞色素P450 2B4中轴向半胱氨酸-436配体被组氨酸取代后，加氧酶功能丧失，NADPH氧化酶活性保留，且出现过氧化物酶活性。
Arch Biochem Biophys. 2005 Feb 1;434(1):128-38. doi: 10.1016/j.abb.2004.10.015.
8
A new understanding on how heme metabolism occurs in heme oxygenase: water-assisted oxo mechanism.血红素加氧酶中血红素代谢发生的新认识：水辅助的氧合机制。
Dalton Trans. 2012 Oct 14;41(38):11642-50. doi: 10.1039/c2dt30777d. Epub 2012 Jul 23.
9
Stabilization and spectroscopic characterization of the dioxygen complex of wild-type cytochrome P4502B4 (CYP2B4) and its distal side E301Q, T302A and proximal side F429H mutants at subzero temperatures.野生型细胞色素P4502B4（CYP2B4）及其远侧E301Q、T302A和近侧F429H突变体的双氧配合物在零下温度下的稳定性及光谱表征。
Biochim Biophys Acta. 2011 Jan;1814(1):69-75. doi: 10.1016/j.bbapap.2010.07.012. Epub 2010 Jul 13.
10
Catalytic mechanism of heme oxygenase through EPR and ENDOR of cryoreduced oxy-heme oxygenase and its Asp 140 mutants.通过低温还原的氧合血红素加氧酶及其天冬氨酸140突变体的电子顺磁共振（EPR）和电子核双共振（ENDOR）研究血红素加氧酶的催化机制
J Am Chem Soc. 2002 Feb 27;124(8):1798-808. doi: 10.1021/ja0122391.
引用本文的文献
1
Oxygen Activation and Radical Transformations in Heme Proteins and Metalloporphyrins.血红素蛋白和金属卟啉中的氧活化和自由基转化。
Chem Rev. 2018 Mar 14;118(5):2491-2553. doi: 10.1021/acs.chemrev.7b00373. Epub 2017 Dec 29.
2
Spectroscopic studies of the cytochrome P450 reaction mechanisms.细胞色素 P450 反应机制的光谱研究。
Biochim Biophys Acta Proteins Proteom. 2018 Jan;1866(1):178-204. doi: 10.1016/j.bbapap.2017.06.021. Epub 2017 Jun 28.
3
Oriented electric fields as future smart reagents in chemistry.取向电场——化学领域未来的智能试剂
Nat Chem. 2016 Nov 22;8(12):1091-1098. doi: 10.1038/nchem.2651.
4
Immersive virtual reality in computational chemistry: Applications to the analysis of QM and MM data.计算化学中的沉浸式虚拟现实：在量子力学（QM）和分子力学（MM）数据分析中的应用。
Int J Quantum Chem. 2016 Nov 15;116(22):1731-1746. doi: 10.1002/qua.25207. Epub 2016 Jul 13.
5
Role of the Proximal Cysteine Hydrogen Bonding Interaction in Cytochrome P450 2B4 Studied by Cryoreduction, Electron Paramagnetic Resonance, and Electron-Nuclear Double Resonance Spectroscopy.通过低温还原、电子顺磁共振和电子-核双共振光谱研究近端半胱氨酸氢键相互作用在细胞色素P450 2B4中的作用
Biochemistry. 2016 Feb 16;55(6):869-83. doi: 10.1021/acs.biochem.5b00744. Epub 2016 Feb 3.
6
F429 Regulation of Tunnels in Cytochrome P450 2B4: A Top Down Study of Multiple Molecular Dynamics Simulations.F429对细胞色素P450 2B4中通道的调控：多分子动力学模拟的自上而下研究
PLoS One. 2015 Sep 28;10(9):e0137075. doi: 10.1371/journal.pone.0137075. eCollection 2015.
7
Structural and functional characterization of a cytochrome P450 2B4 F429H mutant with an axial thiolate-histidine hydrogen bond.具有轴向硫醇盐-组氨酸氢键的细胞色素P450 2B4 F429H突变体的结构与功能表征
Biochemistry. 2014 Aug 12;53(31):5080-91. doi: 10.1021/bi5003794. Epub 2014 Jul 31.
8
Applications of density functional theory to iron-containing molecules of bioinorganic interest.密度泛函理论在生物无机感兴趣的含铁分子中的应用。
Front Chem. 2014 Apr 29;2:14. doi: 10.3389/fchem.2014.00014. eCollection 2014.
9
How Do Perfluorinated Alkanoic Acids Elicit Cytochrome P450 to Catalyze Methane Hydroxylation? An MD and QM/MM Study.全氟链烷酸如何引发细胞色素P450催化甲烷羟基化？一项分子动力学和量子力学/分子力学研究。
RSC Adv. 2013 Mar 7;3(9):2995-3005. doi: 10.1039/C2RA22294A.
10
Experimental documentation of the structural consequences of hydrogen-bonding interactions to the proximal cysteine of a cytochrome P450.实验记录了氢键相互作用对细胞色素 P450 近端半胱氨酸的结构影响。
Angew Chem Int Ed Engl. 2012 Oct 8;51(41):10403-7. doi: 10.1002/anie.201205912. Epub 2012 Sep 11.
本文引用的文献
1
Stabilization and spectroscopic characterization of the dioxygen complex of wild-type cytochrome P4502B4 (CYP2B4) and its distal side E301Q, T302A and proximal side F429H mutants at subzero temperatures.野生型细胞色素P4502B4（CYP2B4）及其远侧E301Q、T302A和近侧F429H突变体的双氧配合物在零下温度下的稳定性及光谱表征。
Biochim Biophys Acta. 2011 Jan;1814(1):69-75. doi: 10.1016/j.bbapap.2010.07.012. Epub 2010 Jul 13.
2
Defining the structural consequences of mechanism-based inactivation of mammalian cytochrome P450 2B4 using resonance Raman spectroscopy.使用共振拉曼光谱技术定义哺乳动物细胞色素 P450 2B4 的基于机制失活的结构后果。
J Am Chem Soc. 2010 Feb 10;132(5):1494-5. doi: 10.1021/ja910276s.
3
Heme oxygenase reveals its strategy for catalyzing three successive oxygenation reactions.血红素加氧酶揭示了其催化三个连续氧合反应的策略。
Acc Chem Res. 2010 Feb 16;43(2):240-7. doi: 10.1021/ar9001685.
4
P450 enzymes: their structure, reactivity, and selectivity-modeled by QM/MM calculations.细胞色素P450酶：其结构、反应性和选择性——通过量子力学/分子力学计算建模
Chem Rev. 2010 Feb 10;110(2):949-1017. doi: 10.1021/cr900121s.
5
Molecular oxygen activation and proton transfer mechanisms in lanosterol 14alpha-demethylase catalysis.羊毛甾醇14α-脱甲基酶催化中的分子氧活化和质子转移机制。
J Phys Chem B. 2009 Jun 11;113(23):8170-82. doi: 10.1021/jp902932p.
6
How is the reactivity of cytochrome P450cam affected by Thr252X mutation? A QM/MM study for X = serine, valine, alanine, glycine.苏氨酸252X突变如何影响细胞色素P450cam的反应活性？针对X为丝氨酸、缬氨酸、丙氨酸、甘氨酸的量子力学/分子力学研究。
J Am Chem Soc. 2009 Apr 8;131(13):4755-63. doi: 10.1021/ja808744k.
7
QM/MM methods for biomolecular systems.用于生物分子系统的量子力学/分子力学方法。
Angew Chem Int Ed Engl. 2009;48(7):1198-229. doi: 10.1002/anie.200802019.
8
Structure and function of an NADPH-cytochrome P450 oxidoreductase in an open conformation capable of reducing cytochrome P450.一种处于开放构象、能够还原细胞色素P450的NADPH-细胞色素P450氧化还原酶的结构与功能
J Biol Chem. 2009 Apr 24;284(17):11374-84. doi: 10.1074/jbc.M807868200. Epub 2009 Jan 26.
9
Compound I in heme thiolate enzymes: a comparative QM/MM study.血红素硫醇盐酶中的化合物I：一项比较性量子力学/分子力学研究。
J Phys Chem A. 2008 Dec 18;112(50):13128-38. doi: 10.1021/jp806770y.
10
Characterization of the microsomal cytochrome P450 2B4 O2 activation intermediates by cryoreduction and electron paramagnetic resonance.通过低温还原和电子顺磁共振对微粒体细胞色素P450 2B4的O2活化中间体进行表征。
Biochemistry. 2008 Sep 9;47(36):9661-6. doi: 10.1021/bi800926x. Epub 2008 Aug 13.
Zotero 插件