通过 35GHz2HENDOR 光谱学确定的甲醛抑制黄嘌呤氧化酶的结构。
The structure of formaldehyde-inhibited xanthine oxidase determined by 35 GHz 2H ENDOR spectroscopy.
机构信息
Chemistry Department, Northwestern University, Evanston, Illinois 60208-3113, USA.
出版信息
J Am Chem Soc. 2010 Oct 13;132(40):14015-7. doi: 10.1021/ja106432h.
Abstract
The formaldehyde-inhibited Mo(V) state of xanthine oxidase (I) has been studied for four decades, yet it has not proven possible to distinguish unequivocally among the several structures proposed for this form. The uniquely large isotropic hyperfine coupling for (13)C from CH(2)O led to the intriguing suggestion of a direct Mo-C bond for the active site of I. This suggestion was supported by the recent crystal structures of glycol- and glycerol-inhibited forms of aldehyde oxidoreductase, a member of the xanthine oxidase family. (1)H and (2)H ENDOR spectra of I(C(1,2)H(2)O) in H(2)O/D(2)O buffer now have unambiguously revealed that the active-site structure of I contains a CH(2)O adduct of Mo(V) in the form of a four-membered ring with S and O linking the C to Mo and have ruled out a direct Mo-C bond. Density functional theory computations are consistent with this conclusion. We interpret the large (13)C coupling as resulting from a "transannular hyperfine interaction".
摘要
黄嘌呤氧化酶（I）的甲醛抑制态已被研究了四十年，但仍未能明确区分为此种形式提出的几种结构。（13）C 与 CH2O 之间独特的各向同性超精细耦合导致了一个有趣的假设，即活性位点的 Mo-C 键。这一假设得到了醛氧化还原酶（黄嘌呤氧化酶家族的一个成员）的乙二醇和甘油抑制形式的最近晶体结构的支持。I（C1,2H2O）在 H2O/D2O 缓冲液中的（1）H 和（2）H ENDOR 光谱现在已经明确揭示，I 的活性位点结构包含 Mo（V）的 CH2O 加合物，形式为具有 S 和 O 连接 C 到 Mo 的四元环，并排除了直接的 Mo-C 键。密度泛函理论计算与这一结论一致。我们将大（13）C 耦合解释为“跨环超精细相互作用”的结果。
相似文献
1
The structure of formaldehyde-inhibited xanthine oxidase determined by 35 GHz 2H ENDOR spectroscopy.
J Am Chem Soc. 2010 Oct 13;132(40):14015-7. doi: 10.1021/ja106432h.
2
35 GHz ENDOR characterization of the "very rapid" signal of xanthine oxidase reacted with 2-hydroxy-6-methylpurine (13C8): evidence against direct Mo-C8 interaction.
J Am Chem Soc. 2001 Mar 21;123(11):2658-63. doi: 10.1021/ja003894w.
3
X-ray crystal structure and EPR spectra of "arsenite-inhibited" Desulfovibriogigas aldehyde dehydrogenase: a member of the xanthine oxidase family.
J Am Chem Soc. 2004 Jul 21;126(28):8614-5. doi: 10.1021/ja0490222.
4
Spectroscopic and electronic structure studies probing covalency contributions to C-H bond activation and transition-state stabilization in xanthine oxidase.
Inorg Chem. 2011 Nov 7;50(21):10919-28. doi: 10.1021/ic201477n. Epub 2011 Oct 5.
5
Evidence favoring molybdenum-carbon bond formation in xanthine oxidase action: 17Q- and 13C-ENDOR and kinetic studies.
Biochemistry. 1996 Feb 6;35(5):1432-43. doi: 10.1021/bi9520500.
6
31P ENDOR studies of xanthine oxidase: coupling of phosphorus of the pterin cofactor to molybdenum (V).
Biochemistry. 1991 Apr 23;30(16):3969-75. doi: 10.1021/bi00230a024.
7
Numbers and exchangeability with water of oxygen-17 atoms coupled to molybdenum (V) in different reduced forms of xanthine oxidase.
Biochemistry. 1982 Nov 9;21(23):5992-9. doi: 10.1021/bi00266a041.
8
Comparative EPR and redox studies of three prokaryotic enzymes of the xanthine oxidase family: quinoline 2-oxidoreductase, quinaldine 4-oxidase, and isoquinoline 1-oxidoreductase.
Biochemistry. 1997 Aug 12;36(32):9780-90. doi: 10.1021/bi970581d.
9
Isotropic exchange interaction between Mo and the proximal FeS center in the xanthine oxidase family member aldehyde oxidoreductase from Desulfovibrio gigas on native and polyalcohol inhibited samples: an EPR and QM/MM study.
J Biol Inorg Chem. 2015 Mar;20(2):233-42. doi: 10.1007/s00775-014-1204-8. Epub 2014 Oct 25.
10
Studies on the mechanism of action of xanthine oxidase.
J Inorg Biochem. 2004 May;98(5):841-8. doi: 10.1016/j.jinorgbio.2003.11.010.
引用本文的文献
1
Magnetic interactions between metal sites in complex enzymes.
J Biol Inorg Chem. 2025 Aug;30(4-5):329-344. doi: 10.1007/s00775-025-02120-1. Epub 2025 Jul 24.
2
Spectroscopic Studies of Mononuclear Molybdenum Enzyme Centers.
Molecules. 2022 Jul 27;27(15):4802. doi: 10.3390/molecules27154802.
3
Molybdenum and Tungsten Cofactors and the Reactions They Catalyze.
Met Ions Life Sci. 2020 Mar 23;20. doi: 10.1515/9783110589757-015.
4
Electronic structure contributions to reactivity in xanthine oxidase family enzymes.
J Biol Inorg Chem. 2015 Mar;20(2):183-94. doi: 10.1007/s00775-014-1212-8. Epub 2014 Nov 26.
5
The mononuclear molybdenum enzymes.
Chem Rev. 2014 Apr 9;114(7):3963-4038. doi: 10.1021/cr400443z. Epub 2014 Jan 28.
6
Orbital contributions to CO oxidation in Mo-Cu carbon monoxide dehydrogenase.
Chem Commun (Camb). 2014 Feb 4;50(9):1104-6. doi: 10.1039/c3cc47705c.
7
(13)C and (63,65)Cu ENDOR studies of CO dehydrogenase from Oligotropha carboxidovorans. Experimental evidence in support of a copper-carbonyl intermediate.
J Am Chem Soc. 2013 Nov 27;135(47):17775-82. doi: 10.1021/ja406136f. Epub 2013 Nov 19.
8
A closer look at the spectroscopic properties of possible reaction intermediates in wild-type and mutant (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase.
Biochemistry. 2012 Jun 19;51(24):4835-49. doi: 10.1021/bi3001215. Epub 2012 Jun 7.
9
Terpene biosynthesis: modularity rules.
Angew Chem Int Ed Engl. 2012 Jan 27;51(5):1124-37. doi: 10.1002/anie.201103110. Epub 2011 Nov 21.
10
Mechanistic studies of an IspH-catalyzed reaction: implications for substrate binding and protonation in the biosynthesis of isoprenoids.
Angew Chem Int Ed Engl. 2011 Dec 16;50(51):12304-7. doi: 10.1002/anie.201104124. Epub 2011 Oct 24.
本文引用的文献
1
Bioorganometallic mechanism of action, and inhibition, of IspH.
Proc Natl Acad Sci U S A. 2010 Mar 9;107(10):4522-7. doi: 10.1073/pnas.0911087107. Epub 2010 Feb 19.
2
Simulating suppression effects in Pulsed ENDOR, and the 'hole in the middle' of Mims and Davies ENDOR Spectra.
Appl Magn Reson. 2010 Jan 1;37(1-4):763-779. doi: 10.1007/s00723-009-0083-6.
3
Formation of {[HIPTN(3)N]Mo(III)H}(-) by heterolytic cleavage of H(2) as established by EPR and ENDOR spectroscopy.
Inorg Chem. 2010 Jan 18;49(2):704-13. doi: 10.1021/ic902006v.
4
ENDOR spectroscopy shows that guanine N1 binds to [4Fe-4S] cluster II of the S-adenosylmethionine-dependent enzyme MoaA: mechanistic implications.
J Am Chem Soc. 2009 Jul 8;131(26):9184-5. doi: 10.1021/ja903978u.
5
Kinetic, structural, and EPR studies reveal that aldehyde oxidoreductase from Desulfovibrio gigas does not need a sulfido ligand for catalysis and give evidence for a direct Mo-C interaction in a biological system.
J Am Chem Soc. 2009 Jun 17;131(23):7990-8. doi: 10.1021/ja809448r.
6
Reductive half-reaction of aldehyde oxidoreductase toward acetaldehyde: a combined QM/MM study.
J Am Chem Soc. 2009 Apr 8;131(13):4628-40. doi: 10.1021/ja805938w.
7
Substrate Orientation and Catalysis at the Molybdenum Site in Xanthine Oxidase: CRYSTAL STRUCTURES IN COMPLEX WITH XANTHINE AND LUMAZINE.
J Biol Chem. 2009 Mar 27;284(13):8760-7. doi: 10.1074/jbc.M804517200. Epub 2008 Dec 24.
8
Substrate orientation in xanthine oxidase: crystal structure of enzyme in reaction with 2-hydroxy-6-methylpurine.
J Biol Chem. 2008 Feb 22;283(8):4818-24. doi: 10.1074/jbc.M707918200. Epub 2007 Dec 6.
9
Oxidation reaction by xanthine oxidase: theoretical study of reaction mechanism.
J Am Chem Soc. 2007 Jul 4;129(26):8131-8. doi: 10.1021/ja068584d. Epub 2007 Jun 12.
10
Probing the N(5)-H bond of the isoalloxazine moiety of flavin radicals by X- and W-band pulsed electron-nuclear double resonance.
Chemphyschem. 2005 Feb;6(2):292-9. doi: 10.1002/cphc.200400377.
Zotero 插件