Excitation of the M intermediates of wild-type bacteriorhodopsin and mutant D96N: Temperature dependence of absorbance, electric responses and proton movements

Rudolf Tóth-Boconádi, Andras Dér, Stefka G. Taneva, Lajos Keszthelyi

Research output: Contribution to journalArticle

Abstract

The simplest proton pump known in biological systems, bacteriorhodopsin (bR), is the first ion-transporting membrane protein, the function of which can be described at the atomic level, with the aid of molecular dynamics calculations. To get additional experimental support for the proposed atomic level description of the function of bR, we studied a quasi-stable state of the protein molecule, the so-called M intermediate that plays a crucial role in the proton pumping process. The temperature dependence of the light-induced events occurring in the photocycle of wild-type bacteriorhodopsin and its mutant D96N were followed in detail. Absorbance changes, electric signals generated by charge motion inside the protein, and movement of protons in the protein solution interface either forward (proton release due to excitation of bR) or backward (uptake of protons due to the M excitation: "back-take") were monitored. The obtained Arrhenius parameters indicate that the proton back-take is triggered by charge rearrangements in the protein similar to the proton release triggered by those during the L → M transition. The time necessary for proton back-take determines the reconstitution time of the bR ground state. The data are expected to be used in theoretical modeling of the bR function. Based on these results, a more detailed photocycle model is established to describe the proton pumping mechanism, implying a formal principle (domino model) that is expected to hold also for other charge transfer proteins.

Original languageEnglish
Pages (from-to)365-373
Number of pages9
JournalTheoretical Chemistry Accounts
Volume125
Issue number3-6
DOIs
Publication statusPublished - Mar 2010

Keywords

  • Buffer effect
  • Charge motion
  • Domino model
  • Orientation
  • Photocycle
  • Proton back-take
  • Purple membrane

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry

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