NMR 研究多铜氧化酶 Fet3p 中三核簇的交换耦合。
NMR study of the exchange coupling in the trinuclear cluster of the multicopper oxidase Fet3p.
机构信息
IBR (Instituto de Biología Molecular y Celular de Rosario), CONICET, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, (S2002LRK), Rosario, Argentina.
出版信息
J Am Chem Soc. 2010 Aug 18;132(32):11191-6. doi: 10.1021/ja1037148.
Abstract
Fet3p from Saccharomyces cerevisiae is a multicopper oxidase (MCO) which oxidizes Fe(2+) to Fe(3+). The electronic structure of the different copper centers in this family of enzymes has been extensively studied and discussed for years with a particular focus on the exchange coupling regime in the trinuclear cluster (TNC). Using NMR spectroscopy we have quantified the exchange coupling constant in the type 3 center in a fully metalated oxidase; this value in Fet3p is significantly higher than that reported for proteins containing isolated type 3 centers as tyrosinase. We also provide evidence of exchange coupling between the type 2 and the type 3 Cu(2+) ions, which supports the crystallographic evidence of dioxygen binding to the TNC. This work provides the foundation for the application of NMR to these complex systems.
摘要
酿酒酵母的 Fet3p 是一种多铜氧化酶（MCO），能将 Fe(2+)氧化为 Fe(3+)。多年来，人们对该酶家族中不同铜中心的电子结构进行了广泛的研究和讨论，特别关注三核簇（TNC）中的交换耦合机制。我们利用 NMR 光谱技术，在完全金属化的氧化酶中定量了 3 型中心的交换耦合常数；该值在 Fet3p 中明显高于含有酪氨酸酶等分离型 3 型中心的蛋白质。我们还提供了证据表明 2 型和 3 型 Cu(2+)离子之间存在交换耦合，这支持了 TNC 与氧结合的晶体学证据。这项工作为将 NMR 应用于这些复杂体系奠定了基础。
相似文献
1
NMR study of the exchange coupling in the trinuclear cluster of the multicopper oxidase Fet3p.NMR 研究多铜氧化酶 Fet3p 中三核簇的交换耦合。
J Am Chem Soc. 2010 Aug 18;132(32):11191-6. doi: 10.1021/ja1037148.
2
Spectroscopic characterization and O2 reactivity of the trinuclear Cu cluster of mutants of the multicopper oxidase Fet3p.多铜氧化酶Fet3p突变体的三核铜簇的光谱表征及O2反应活性
Biochemistry. 2002 May 21;41(20):6438-48. doi: 10.1021/bi011979j.
3
Spectroscopic analysis of the trinuclear cluster in the Fet3 protein from yeast, a multinuclear copper oxidase.对来自酵母的Fet3蛋白（一种多核铜氧化酶）中的三核簇进行光谱分析。
Biochemistry. 2000 Mar 7;39(9):2316-24. doi: 10.1021/bi992334a.
4
Role of aspartate 94 in the decay of the peroxide intermediate in the multicopper oxidase Fet3p.天冬氨酸94在多铜氧化酶Fet3p中过氧化物中间体衰变中的作用。
Biochemistry. 2005 Apr 26;44(16):6081-91. doi: 10.1021/bi047379c.
5
Ferrous binding to the multicopper oxidases Saccharomyces cerevisiae Fet3p and human ceruloplasmin: contributions to ferroxidase activity.亚铁与多铜氧化酶酿酒酵母Fet3p和人铜蓝蛋白的结合：对铁氧化酶活性的贡献。
J Am Chem Soc. 2004 Jun 2;126(21):6579-89. doi: 10.1021/ja049220t.
6
Spectroscopy and reactivity of the type 1 copper site in Fet3p from Saccharomyces cerevisiae: correlation of structure with reactivity in the multicopper oxidases.酿酒酵母Fet3p中1型铜位点的光谱学与反应活性：多铜氧化酶中结构与反应活性的相关性
J Am Chem Soc. 2001 Jun 13;123(23):5507-17. doi: 10.1021/ja003975s.
7
Spectroscopic studies of perturbed T1 Cu sites in the multicopper oxidases Saccharomyces cerevisiae Fet3p and Rhus vernicifera laccase: allosteric coupling between the T1 and trinuclear Cu sites.酿酒酵母Fet3p和漆树漆酶中受扰T1铜位点的光谱研究：T1与三核铜位点之间的变构偶联
Biochemistry. 2008 Feb 19;47(7):2036-45. doi: 10.1021/bi7020052. Epub 2008 Jan 16.
8
In vitro unfolding of yeast multicopper oxidase Fet3p variants reveals unique role of each metal site.酵母多铜氧化酶Fet3p变体的体外解折叠揭示了每个金属位点的独特作用。
Proc Natl Acad Sci U S A. 2008 Dec 9;105(49):19258-63. doi: 10.1073/pnas.0806431105. Epub 2008 Nov 25.
9
Structural basis of the ferrous iron specificity of the yeast ferroxidase, Fet3p.酵母铁氧化酶Fet3p对亚铁离子特异性的结构基础
Biochemistry. 2006 Oct 24;45(42):12741-9. doi: 10.1021/bi061543+.
10
Targeted suppression of the ferroxidase and iron trafficking activities of the multicopper oxidase Fet3p from Saccharomyces cerevisiae.对酿酒酵母多铜氧化酶Fet3p的铁氧化酶和铁转运活性进行靶向抑制。
J Biol Inorg Chem. 2003 Jul;8(6):611-20. doi: 10.1007/s00775-003-0456-5. Epub 2003 Apr 9.
引用本文的文献
1
Towards resolving the complex paramagnetic nuclear magnetic resonance (NMR) spectrum of small laccase: assignments of resonances to residue-specific nuclei.解析小型漆酶复杂的顺磁核磁共振（NMR）谱：将共振峰归属到特定残基的原子核
Magn Reson (Gott). 2021 Jan 29;2(1):15-23. doi: 10.5194/mr-2-15-2021. eCollection 2021.
2
Chemical Exchange at the Trinuclear Copper Center of Small Laccase from Streptomyces coelicolor.天蓝色链霉菌小漆酶三核铜中心的化学交换
Biophys J. 2020 Jul 7;119(1):9-14. doi: 10.1016/j.bpj.2020.05.022. Epub 2020 May 29.
3
Rotating magnetic field as tool for enhancing enzymes properties - laccase case study.旋转磁场作为增强酶特性的工具——漆酶案例研究。
Sci Rep. 2019 Mar 6;9(1):3707. doi: 10.1038/s41598-019-39198-y.
4
The copper regulon of the human fungal pathogen Cryptococcus neoformans H99.人类真菌病原体新型隐球菌 H99 的铜调控基因簇。
Mol Microbiol. 2011 Sep;81(6):1560-76. doi: 10.1111/j.1365-2958.2011.07794.x. Epub 2011 Aug 23.
本文引用的文献
1
Multicopper oxidases: a workshop on copper coordination chemistry, electron transfer, and metallophysiology.多铜氧化酶：关于铜配位化学、电子转移和金属生理学的研讨会。
J Biol Inorg Chem. 2010 Jan;15(1):15-28. doi: 10.1007/s00775-009-0590-9. Epub 2009 Oct 9.
2
Geometric and electronic structure differences between the type 3 copper sites of the multicopper oxidases and hemocyanin/tyrosinase.多铜氧化酶与血蓝蛋白/酪氨酸酶的3型铜位点之间的几何结构和电子结构差异。
Proc Natl Acad Sci U S A. 2009 Apr 21;106(16):6585-90. doi: 10.1073/pnas.0902127106. Epub 2009 Apr 3.
3
X-ray structure of a two-domain type laccase: a missing link in the evolution of multi-copper proteins.双结构域I型漆酶的X射线结构：多铜蛋白进化中的缺失环节。
FEBS Lett. 2009 Apr 2;583(7):1189-95. doi: 10.1016/j.febslet.2009.03.008. Epub 2009 Mar 11.
4
Electronic structure of the ground and excited states of the Cu(A) site by NMR spectroscopy.通过核磁共振光谱法研究铜（A）位点基态和激发态的电子结构。
J Am Chem Soc. 2009 Feb 11;131(5):1939-46. doi: 10.1021/ja8079669.
5
The structure of the small laccase from Streptomyces coelicolor reveals a link between laccases and nitrite reductases.来自天蓝色链霉菌的小型漆酶结构揭示了漆酶与亚硝酸还原酶之间的联系。
J Mol Biol. 2009 Jan 30;385(4):1165-78. doi: 10.1016/j.jmb.2008.11.024. Epub 2008 Nov 25.
6
O2 reduction to H2O by the multicopper oxidases.多铜氧化酶将氧气还原为水。
Dalton Trans. 2008 Aug 14(30):3921-32. doi: 10.1039/b800799c. Epub 2008 May 7.
7
Direct identification of a bacterial manganese(II) oxidase, the multicopper oxidase MnxG, from spores of several different marine Bacillus species.从几种不同海洋芽孢杆菌的孢子中直接鉴定出一种细菌锰（II）氧化酶——多铜氧化酶MnxG。
Appl Environ Microbiol. 2008 Mar;74(5):1527-34. doi: 10.1128/AEM.01240-07. Epub 2007 Dec 28.
8
Electronic structure of the peroxy intermediate and its correlation to the native intermediate in the multicopper oxidases: insights into the reductive cleavage of the o-o bond.多铜氧化酶中过氧中间体的电子结构及其与天然中间体的相关性：对O-O键还原裂解的见解
J Am Chem Soc. 2007 Oct 31;129(43):13127-36. doi: 10.1021/ja073947a. Epub 2007 Oct 5.
9
Spectroscopic and kinetic studies of perturbed trinuclear copper clusters: the role of protons in reductive cleavage of the O-O bond in the multicopper oxidase Fet3p.受扰三核铜簇的光谱学和动力学研究：质子在多铜氧化酶Fet3p中O-O键还原断裂中的作用
J Am Chem Soc. 2007 Oct 31;129(43):13118-26. doi: 10.1021/ja073905m. Epub 2007 Oct 5.
10
Crystal structure of a blue laccase from Lentinus tigrinus: evidences for intermediates in the molecular oxygen reductive splitting by multicopper oxidases.虎皮香菇蓝色漆酶的晶体结构：多铜氧化酶分子氧还原分裂中间体的证据
BMC Struct Biol. 2007 Sep 26;7:60. doi: 10.1186/1472-6807-7-60.
Zotero 插件