Identifying the mechanism of second proton release from the bc1 complex Qo-site

Wilson, Charles Alexander

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https://hdl.handle.net/2142/97425 Copy

Description

Title

Identifying the mechanism of second proton release from the bc1 complex Qo-site

Author(s)

Wilson, Charles Alexander

Issue Date

2017-04-21

Director of Research (if dissertation) or Advisor (if thesis)

Crofts, Antony

Doctoral Committee Chair(s)

Crofts, Antony

Committee Member(s)

• Aksimentiev, Oleksii
• Gennis, Robert
• Gruebele, Martin

Department of Study

School of Molecular & Cell Bio

Discipline

Biophysics & Computnl Biology

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

• Bc1 complex
• Rhodobacter sphaeroides

Abstract

The cytochrome bc1 complex and its family members play a central role in biological energy transduction across all domains of life, oxidizing QH2 in a bifurcated reaction which generates a proton-motive force used for ATP synthesis. It is widely accepted to operate according to a modified Q-cycle, but there is still debate over some aspects of the mechanism. We used the pH-sensitive dye neutral red to study the release of protons from the bc1 complex, with specific focus of the mechanism for release of the second proton from the Qo-site. Partial processes for the proton release could be isolated through use of different inhibitors, and correlated with electron transfers through the complex. We tested mutations at residues R94, N279, and Y147 in the b subunit of the complex, which are thought to interact with the water chain which provides a pathway for release of the second proton. Mutants R94A, N279F, and Y147T showed a significant decrease in proton release for a second turnover compared to the first, an indicator that the proton from the first turnover had difficulty leaving the complex. The latter two mutants also showed an intriguing difference in kinetics for the proton release and electron transfers for the first turnover.

Graduation Semester

2017-05

Type of Resource

text

Permalink

http://hdl.handle.net/2142/97425

Copyright and License Information

Copyright 2017 Charles Wilson

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