Multiconformation Continuum Electrostatics Analysis of the Effects of a Buried Asp Introduced near Heme a in Rhodobacter sphaeroides Cytochrome c Oxidase
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- 作者：Jun Zhang ; M. R. Gunner
- 刊名：Biochemistry
- 出版年：2010
- 出版时间：September 21, 2010
- 年：2010
- 卷：49
- 期：37
- 页码：8043-8052
- 全文大小：938K
- 年卷期：v.49,no.37(September 21, 2010)
- ISSN：1520-4995
文摘
Cytochrome c oxidase (CcO) reduces O2 to water via a series of proton-coupled electron transfers, generating a transmembrane electrochemical gradient. Coupling electron and proton transfer requires changing the pKa values of buried residues at each stage in the reaction cycle. Heme a is a key cofactor in the CcO electron transfer chain. Mutation of Ser44 to Asp has been reported [Mills, D. A., et al. (2008) Biochemistry 47, 11499−11509], changing the hydrogen bond acceptor from His102, the heme a axial ligand in Rhodobactor sphaeroides CcO. This adds an acidic residue to the CcO interior. The electrochemical behavior of heme a in wild-type and S44D CcO is compared using the continuum electrostatics program MCCE. The introduced, deeply buried Asp remains ionized at physiological pH only when the nearby heme is oxidized. Heme a reduction is now calculated to be strongly coupled to Asp proton binding, while with Ser44, it is weakly coupled to small protonation shifts at multiple sites, increasing the pH dependence in the mutant. At pH 7, the partially ionized Asp 44 is calculated to lower the heme redox potential by 50 mV as expected given the thermodynamics of coupled electron and proton transfers. This highlights an curious finding in the experimental results where a low Asp pKa is found together with a stabilized reduced heme. The stabilization of a heme oxidation in a model complex by a hydrogen bond to the axial His ligand calculated with continuum electrostatics and with density functional theory were in good agreement.