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Recombinant Expression, Purification, and Reconstitution of the Chloroplast ATP Synthase c-subunit Ring


Abstract ATP synthase is a large multimeric protein complex responsible for generating the energy molecule adenosine triphosphate (ATP) in most organisms. The catalysis involves the rotation of a ring of c-subunits, which is driven by the transmembrane electrochemical gradient. This dissertation reports how the eukaryotic c-subunit from spinach chloroplast ATP synthase has successfully been expressed in Escherichia coli and purified in mg quantities by incorporating a unique combination of methods. Expression was accomplished using a codon optimized gene for the c-subunit, and it was expressed as an attachment to the larger, more soluble, native maltose binding protein (MBP-c1). The fusion protein MBP-c1 was purified on an affinity column, and the ... (more)
Created Date 2011
Contributor Lawrence, Robert Michael (Author) / Fromme, Petra (Advisor) / Chen, Julian J.L. (Committee member) / Woodbury, Neal W. (Committee member) / Arizona State University (Publisher)
Subject Biochemistry / Biophysics / Biology, Plant Physiology / ATP synthase / chloroplast / coupling ratio / recombinant expression / ring stoichiometry / subunit c III ring
Type Doctoral Dissertation
Extent 215 pages
Language English
Copyright
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Note Ph.D. Biochemistry 2011
Collaborating Institutions Graduate College / ASU Library
Additional Formats MODS / OAI Dublin Core / RIS


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Description Dissertation/Thesis