Retardation of proton transfer caused by binding of the transition metal ion to the bacterial reaction center is due to pKa shifts of key protonatable residues

L. Gerencsér, P. Maróti

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Transition metal ions bind to the reaction center (RC) protein of the photosynthetic bacterium Rhodobacter sphaeroides and slow the light-induced electron and proton transfer to the secondary quinone, QB. We studied the properties of the metal ion-RC complex by measuring the pH dependence of the dissociation constant and the stoichiometry of proton release upon ligand formation. We investigated the mechanism of inhibition by measuring the stoichiometry and kinetics of flash-induced proton binding, the transfer of (first and second) electrons to QB, and the rate of steady-state turnover of the RC in the absence and presence of Cd2+ and Ni2+ on a wide pH range. The following results were obtained. (1) The complexation of transition metal ions Cd2+ and Ni2+ with the bacterial RC showed strong pH dependence. This observation was explained by different (pH-dependent) states of the metal-ligand cluster: the complex formation was strong when the ligand (Asp and His residues) was deprotonated and was much weaker if the ligand was partly (or fully) protonated. A direct consequence of the model was the pH-dependent proton release upon complexation. (2) The retardation of transfer of electrons and protons to QB was also strongly pH-dependent. The effect was large in the neutral pH range and decreased toward the acidic and alkaline pH values. (3) Steady-state turnover measurements indicated that the rate of the second proton transfer was much less inhibited than that of the first one, which became the rate-limiting step in continuous turnover of the RC. (4) Sodium azide partly recovered the proton transfer rate. The effect is not due to removal of the bound metal ion by azide but probably by formation of a proton-transporting azide network similarly as water molecules may build up proton pathways. (5) We argue that the inhibition comes mainly from pKa shifts of key protonatable residues that control the proton transfer along the H-bond network to QB. The electrostatic interaction between the metal ion and these residues may result in acidic pKa shifts between 1.5 and 2.0 that account for the observed retardation of the electron and proton transfer.

Original languageEnglish
Pages (from-to)1850-1860
Number of pages11
JournalBiochemistry
Volume40
Issue number6
DOIs
Publication statusPublished - Feb 13 2001

ASJC Scopus subject areas

  • Biochemistry

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