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- Creators: Arizona State University
- Creators: Alford, Eddie J
Description
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 c1 subunit was subsequently severed by protease cleavage in the presence of detergent. Final purification of the monomeric c1 subunit was accomplished using reversed phase column chromatography with ethanol as an eluent. Circular dichroism spectroscopy data showed clear evidence that the purified c-subunit is folded with the native alpha-helical secondary structure. Recent experiments appear to indicate that this monomeric recombinant c-subunit forms an oligomeric ring that is similar to its native tetradecameric form when reconstituted in liposomes. The F-type ATP synthase c-subunit stoichiometry is currently known to vary from 8 to 15 subunits among different organisms. This has a direct influence on the metabolic requirements of the corresponding organism because each c-subunit binds and transports one H+ across the membrane as the ring makes a complete rotation. The c-ring rotation drives rotation of the gamma-subunit, which in turn drives the synthesis of 3 ATP for every complete rotation. The availability of a recombinantly produced c-ring will lead to new experiments which can be designed to investigate the possible factors that determine the variable c-ring stoichiometry and structure.
ContributorsLawrence, Robert Michael (Author) / Fromme, Petra (Thesis advisor) / Chen, Julian J.L. (Committee member) / Woodbury, Neal W. (Committee member) / Arizona State University (Publisher)
Created2011
Description
This study was designed to produce a comprehensive flora of Usery Mountain Regional Park and Pass Mountain of the Tonto National Forest. A total of 168 vascular plant species representing 46 families and 127 genera were collected or documented at this study area. Sixteen species were not native to the flora of Arizona and represent 9.5% of the flora. Nevertheless, the study area does not appear to be significantly damaged or degraded in spite of its historical and current land use. The location and types of invasive species recorded in this study will assist with implementing preventative measures to prevent further spreading of certain species. The complete list of all vascular species recorded in this study will provide a valuable tool for land management decisions and future restoration projects that may occur at this area or similar sites and invasive species control. The distribution of the saguaro (Carnegiea gigantea) population on Pass Mountain was documented through the measurement of saguaros by random sampling. ArcGIS was used to generate 50 random points for sampling the saguaro population. Analysis to determine saguaro habitat preferences based on the parameters of aspect, slope and elevation was conducted through ArcGIS. The saguaro population of Pass Mountain significantly favored the southern aspects with the highest concentration occurring in the southwest aspects at an average density of 42.66 saguaros per hectare. The large numbers of saguaros recorded in the younger size classes suggests a growing populations.
ContributorsMarshall, Laura Lee (Author) / Steele, Kelly P (Thesis advisor) / Miller, William H. (Committee member) / Alford, Eddie J (Committee member) / Arizona State University (Publisher)
Created2011
Description
The Arizona State University Herbarium began in 1896 when Professor Fredrick Irish collected the first recorded Arizona specimen for what was then called the Tempe Normal School - a Parkinsonia microphylla. Since then, the collection has grown to approximately 400,000 specimens of vascular plants and lichens. The most recent project includes the digitization - both the imaging and databasing - of approximately 55,000 vascular plant specimens from Latin America. To accomplish this efficiently, possibilities in non-traditional methods, including both new and existing technologies, were explored. SALIX (semi-automatic label information extraction) was developed as the central tool to handle automatic parsing, along with BarcodeRenamer (BCR) to automate image file renaming by barcode. These two developments, combined with existing technologies, make up the SALIX Method. The SALIX Method provides a way to digitize herbarium specimens more efficiently than the traditional approach of entering data solely through keystroking. Using digital imaging, optical character recognition, and automatic parsing, I found that the SALIX Method processes data at an average rate that is 30% faster than typing. Data entry speed is dependent on user proficiency, label quality, and to a lesser degree, label length. This method is used to capture full specimen records, including close-up images where applicable. Access to biodiversity data is limited by the time and resources required to digitize, but I have found that it is possible to do so at a rate that is faster than typing. Finally, I experiment with the use of digital field guides in advancing access to biodiversity data, to stimulate public engagement in natural history collections.
ContributorsBarber, Anne Christine (Author) / Landrum, Leslie R. (Thesis advisor) / Wojciechowski, Martin F. (Thesis advisor) / Gilbert, Edward (Committee member) / Lafferty, Daryl (Committee member) / Arizona State University (Publisher)
Created2012