Thomas M Duncan, Ph.D. Associate Professor, Biochemistry and Molecular Biology Associate Professor, Biochemistry and Molecular Biology 4219 Weiskotten Hall Upstate Medical University 750 East Adams Street Syracuse, NY 13210 (315) 464-8711

Education and Clinical Training

Research Program and Department Affiliations

Research Interests

Research Abstract

My research focuses on the structure and function of the ATP synthase - an energy-coupling enzyme complex that is critical to the energy metabolism of most living things. The ATP synthase is located on the inner cell membrane of bacteria, the inner mitochondrial membrane of eukaryotes and on the thylakoid membrane of plant chloroplasts. In the terminal step of oxidative- and photo-phosphorylation, it extracts energy from a transmembrane, electrochemical gradient of protons to drive the synthesis of most cellular ATP.

Background

The ATP synthase consists of two sub-complexes with distinct but coupled functions. The F₀complex spans the membrane bilayer and transports protons. F₁ is a peripheral complex that extends into the aqueous phase and contains three catalytic nucleotide binding sites for ATP synthesis (one visible in cartoon). F₀ and F₁ are coupled through two stalk-like connections of subunits, a central rotor shaft and a peripheral stator. My earlier studies with Richard Cross provided key initial evidence that energy coupling by the ATP synthase involves

1. rotation of the central shaft within F₁ to coordinate the actions of three cooperative, alternating catalytic nucleotide sites, and
2. rotation of subunits within F₀ during energy-driven proton transport.

Remarkably, energy coupling by the ATP synthase is very efficient and reversible: respiration-driven proton transport through F₀ can drive net synthesis of ATP, but net hydrolysis of ATP can also be used to drive proton transport in the opposite direction, thereby generating a transmembrane, electrochemical proton gradient.

Experimental system

Our research primarily uses the bacterial ATP synthase from Escherichia coli, which provides a simple yet powerful model system for structure/function studies of this rotary motor enzyme. The E. coli system provides straightforward genetic screening and engineering of the ATP synthase, and also allows large-scale purification of F₀F₁ and isolated F₁ for biochemical studies.

Current projects

1. Determine high-resolution structures of E. coli F₁ and F₀F₁ by X-ray crystallography. A collaboration with Dr. Gino Cingolani, an experienced protein crystallographer in our department.
2. Determine how the rotor-associated epsilon subunit regulates activity of E. coli F₁, and how epsilon may respond to changes in the electrochemical proton gradient to regulate the activty of membrane-bound E. coli F₀F₁.
3. Engineer specific cysteines into different subunits of F₀F₁ for fluorescent labeling and assays of functional subunit rotation by single-particle fluorescence resonance energy transfer (sp-FRET). A preliminary collaboration with single-molecule microscopist, Dr. Michael B¬örsch (Physics Institute, University of Stuttgart, Germany).

Publications - link to PubMed

Additional Information

This profile was last updated on 10/16/2009

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http://www.upstate.edu/biochem/faculty.php?ID=duncant

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