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This thesis explores a wide array of topics related to the protein folding problem, ranging from the folding mechanism, ab initio structure prediction and protein design, to the mechanism of protein functional evolution, using multi-scale approaches. To investigate the role of native topology on folding mechanism, the native topology is

This thesis explores a wide array of topics related to the protein folding problem, ranging from the folding mechanism, ab initio structure prediction and protein design, to the mechanism of protein functional evolution, using multi-scale approaches. To investigate the role of native topology on folding mechanism, the native topology is dissected into non-local and local contacts. The number of non-local contacts and non-local contact orders are both negatively correlated with folding rates, suggesting that the non-local contacts dominate the barrier-crossing process. However, local contact orders show positive correlation with folding rates, indicating the role of a diffusive search in the denatured basin. Additionally, the folding rate distribution of E. coli and Yeast proteomes are predicted from native topology. The distribution is fitted well by a diffusion-drift population model and also directly compared with experimentally measured half life. The results indicate that proteome folding kinetics is limited by protein half life. The crucial role of local contacts in protein folding is further explored by the simulations of WW domains using Zipping and Assembly Method. The correct formation of N-terminal β-turn turns out important for the folding of WW domains. A classification model based on contact probabilities of five critical local contacts is constructed to predict the foldability of WW domains with 81% accuracy. By introducing mutations to stabilize those critical local contacts, a new protein design approach is developed to re-design the unfoldable WW domains and make them foldable. After folding, proteins exhibit inherent conformational dynamics to be functional. Using molecular dynamics simulations in conjunction with Perturbation Response Scanning, it is demonstrated that the divergence of functions can occur through the modification of conformational dynamics within existing fold for β-lactmases and GFP-like proteins: i) the modern TEM-1 lactamase shows a comparatively rigid active-site region, likely reflecting adaptation for efficient degradation of a specific substrate, while the resurrected ancient lactamases indicate enhanced active-site flexibility, which likely allows for the binding and subsequent degradation of different antibiotic molecules; ii) the chromophore and attached peptides of photocoversion-competent GFP-like protein exhibits higher flexibility than the photocoversion-incompetent one, consistent with the evolution of photocoversion capacity.
ContributorsZou, Taisong (Author) / Ozkan, Sefika B (Thesis advisor) / Thorpe, Michael F (Committee member) / Woodbury, Neal W (Committee member) / Vaiana, Sara M (Committee member) / Ghirlanda, Giovanna (Committee member) / Arizona State University (Publisher)
Created2014
Description
Despite the wealth of folk music traditions in Portugal and the importance of the clarinet in the music of bandas filarmonicas, it is uncommon to find works featuring the clarinet using Portuguese folk music elements. In the interest of expanding this type of repertoire, three new works were commissioned from

Despite the wealth of folk music traditions in Portugal and the importance of the clarinet in the music of bandas filarmonicas, it is uncommon to find works featuring the clarinet using Portuguese folk music elements. In the interest of expanding this type of repertoire, three new works were commissioned from three different composers. The resulting works are Seres Imaginarios 3 by Luis Cardoso; Delirio Barroco by Tiago Derrica; and Memória by Pedro Faria Gomes. In an effort to submit these new works for inclusion into mainstream performance literature, the author has recorded these works on compact disc. This document includes interview transcripts with each composer, providing first-person discussion of each composition, as well as detailed biographical information on each composer. To provide context, the author has included a brief discussion on Portuguese folk music, and in particular, the role that the clarinet plays in Portuguese folk music culture.
ContributorsFerreira, Wesley (Contributor) / Spring, Robert S (Thesis advisor) / Bailey, Wayne (Committee member) / Gardner, Joshua (Committee member) / Hill, Gary (Committee member) / Schuring, Martin (Committee member) / Solis, Theodore (Committee member) / Arizona State University (Publisher)
Created2013
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Description
Proteins are a fundamental unit in biology. Although proteins have been extensively studied, there is still much to investigate. The mechanism by which proteins fold into their native state, how evolution shapes structural dynamics, and the dynamic mechanisms of many diseases are not well understood. In this thesis, protein folding

Proteins are a fundamental unit in biology. Although proteins have been extensively studied, there is still much to investigate. The mechanism by which proteins fold into their native state, how evolution shapes structural dynamics, and the dynamic mechanisms of many diseases are not well understood. In this thesis, protein folding is explored using a multi-scale modeling method including (i) geometric constraint based simulations that efficiently search for native like topologies and (ii) reservoir replica exchange molecular dynamics, which identify the low free energy structures and refines these structures toward the native conformation. A test set of eight proteins and three ancestral steroid receptor proteins are folded to 2.7Å all-atom RMSD from their experimental crystal structures. Protein evolution and disease associated mutations (DAMs) are most commonly studied by in silico multiple sequence alignment methods. Here, however, the structural dynamics are incorporated to give insight into the evolution of three ancestral proteins and the mechanism of several diseases in human ferritin protein. The differences in conformational dynamics of these evolutionary related, functionally diverged ancestral steroid receptor proteins are investigated by obtaining the most collective motion through essential dynamics. Strikingly, this analysis shows that evolutionary diverged proteins of the same family do not share the same dynamic subspace. Rather, those sharing the same function are simultaneously clustered together and distant from those functionally diverged homologs. This dynamics analysis also identifies 77% of mutations (functional and permissive) necessary to evolve new function. In silico methods for prediction of DAMs rely on differences in evolution rate due to purifying selection and therefore the accuracy of DAM prediction decreases at fast and slow evolvable sites. Here, we investigate structural dynamics through computing the contribution of each residue to the biologically relevant fluctuations and from this define a metric: the dynamic stability index (DSI). Using DSI we study the mechanism for three diseases observed in the human ferritin protein. The T30I and R40G DAMs show a loss of dynamic stability at the C-terminus helix and nearby regulatory loop, agreeing with experimental results implicating the same regulatory loop as a cause in cataracts syndrome.
ContributorsGlembo, Tyler J (Author) / Ozkan, Sefika B (Thesis advisor) / Thorpe, Michael F (Committee member) / Ros, Robert (Committee member) / Kumar, Sudhir (Committee member) / Shumway, John (Committee member) / Arizona State University (Publisher)
Created2011
ContributorsBurton, Charlotte (Performer) / ASU Library. Music Library (Publisher)
Created2018-04-08
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Description
Infections caused by the Hepatitis C Virus (HCV) are very common worldwide, affecting up to 3% of the population. Chronic infection of HCV may develop into liver cirrhosis and liver cancer which is among the top five of the most common cancers. Therefore, vaccines against HCV are under intense study

Infections caused by the Hepatitis C Virus (HCV) are very common worldwide, affecting up to 3% of the population. Chronic infection of HCV may develop into liver cirrhosis and liver cancer which is among the top five of the most common cancers. Therefore, vaccines against HCV are under intense study in order to prevent HCV from harming people's health. The envelope protein 2 (E2) of HCV is thought to be a promising vaccine candidate because it can directly bind to a human cell receptor and plays a role in viral entry. However, the E2 protein production in cells is inefficient due to its complicated matured structure. Folding of E2 in the endoplasmic reticulum (ER) is often error-prone, resulting in production of aggregates and misfolded proteins. These incorrect forms of E2 are not functional because they are not able to bind to human cells and stimulate antibody response to inhibit this binding. This study is aimed to overcome the difficulties of HCV E2 production in plant system. Protein folding in the ER requires great assistance from molecular chaperones. Thus, in this study, two molecular chaperones in the ER, calreticulin and calnexin, were transiently overexpressed in plant leaves in order to facilitate E2 folding and production. Both of them showed benefits in increasing the yield of E2 and improving the quality of E2. In addition, poorly folded E2 accumulated in the ER may cause stress in the ER and trigger transcriptional activation of ER molecular chaperones. Therefore, a transcription factor involved in this pathway, named bZIP60, was also overexpressed in plant leaves, aiming at up-regulating a major family of molecular chaperones called BiP to assist protein folding. However, our results showed that BiP mRNA levels were not up-regulated by bZIP60, but they increased in response to E2 expression. The Western blot analysis also showed that overexpression of bZIP60 had a small effect on promoting E2 folding. Overall, this study suggested that increasing the level of specific ER molecular chaperones was an effective way to promote HCV E2 protein production and maturation.
ContributorsHong, Fan (Author) / Mason, Hugh (Thesis advisor) / Gaxiola, Roberto (Committee member) / Chang, Yung (Committee member) / Chen, Qiang (Committee member) / Arizona State University (Publisher)
Created2011
ContributorsDruesedow, Elizabeth (Performer) / ASU Library. Music Library (Publisher)
Created2018-04-07
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Description
Quercetin 2,3-dioxygenase from Bacillus subtilis has been identified and characterized as the first known prokaryotic quercetinase. This enzyme catalyzes the cleavage of the O-heteroaromatic ring of the flavonol quercetin to the corresponding depside and carbon monoxide. The first quercetinase was characterized from a species of Aspergillus genus, and was found

Quercetin 2,3-dioxygenase from Bacillus subtilis has been identified and characterized as the first known prokaryotic quercetinase. This enzyme catalyzes the cleavage of the O-heteroaromatic ring of the flavonol quercetin to the corresponding depside and carbon monoxide. The first quercetinase was characterized from a species of Aspergillus genus, and was found to contain one Cu2+ per subunit. For many years, it was thought that the B. subtilis quercetinase contained two Fe2+ ions per subunit; however, it has since been discovered that Mn2+ is a much more likely cofactor. Studies of overexpressed bacterial enzyme in E. coli indicated that this enzyme may be active with other metal ions (e.g. Co2+); however, the production of enzyme with full metal incorporation has only been possible with Mn2+. This study explores the notion that metal manipulation after translation, by partially unfolding the enzyme, chelating the metal ions, and then refolding the protein in the presence of an excess of divalent metal ions, could generate enzyme with full metal occupancy. The protocols presented here included testing for activity after incubating purified quercetinase with EDTA, DDTC, imidazole and GndHCl. It was found that the metal chelators had little to no effect on quercetinase activity. Imidazole did appear to inhibit the enzyme at concentrations in the millimolar range. In addition, the quercetinase was denatured in GndHCl at concentrations above 1 M. Recovering an active enzyme after partial or complete unfolding proved difficult, if not impossible.
ContributorsKrojanker, Elan Daniel (Author) / Francisco, Wilson (Thesis director) / Allen, James P. (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05
Description
This project includes a recording and performance guide for three newly commissioned pieces for the clarinet. The first piece, shimmer, was written by Grant Jahn and is for B-flat clarinet and electronics. The second piece, Paragon, is for B-flat clarinet and piano and was composed by Dr. Theresa Martin. The

This project includes a recording and performance guide for three newly commissioned pieces for the clarinet. The first piece, shimmer, was written by Grant Jahn and is for B-flat clarinet and electronics. The second piece, Paragon, is for B-flat clarinet and piano and was composed by Dr. Theresa Martin. The third and final piece, Duality in the Eye of a Bovine, was written by Kurt Mehlenbacher and is for B-flat clarinet, bass clarinet, and piano. In addition to the performance guide, this document also includes background information and program notes for the compositions, as well as composer biographical information, a list of other works featuring the clarinet by each composer, and transcripts of composer and performer interviews. This document is accompanied by a recording of the three pieces.
ContributorsPoupard, Caitlin Marie (Author) / Spring, Robert (Thesis advisor) / Gardner, Joshua (Thesis advisor) / Hill, Gary (Committee member) / Oldani, Robert (Committee member) / Schuring, Martin (Committee member) / Arizona State University (Publisher)
Created2016
Description
The primary objective of this research project is to expand the clarinet repertoire with the addition of four new pieces. Each of these new pieces use contemporary clarinet techniques, including electronics, prerecorded sounds, multiphonics, circular breathing, multiple articulation, demi-clarinet, and the clari-flute. The repertoire composed includes Grant Jahn’s Duo for

The primary objective of this research project is to expand the clarinet repertoire with the addition of four new pieces. Each of these new pieces use contemporary clarinet techniques, including electronics, prerecorded sounds, multiphonics, circular breathing, multiple articulation, demi-clarinet, and the clari-flute. The repertoire composed includes Grant Jahn’s Duo for Two Clarinets, Reggie Berg’s Funkalicious for Clarinet and Piano, Rusty Banks’ Star Juice for Clarinet and Fixed Media, and Chris Malloy’s A Celestial Breath for Clarinet and Electronics. In addition to the musical commissions, this project also includes interviews with the composers indicating how they wrote these works and what their influences were, along with any information pertinent to the performer, professional recordings of each piece, as well as performance notes and suggestions.
ContributorsCase-Ruchala, Celeste Ann (Contributor) / Gardner, Joshua (Thesis advisor) / Spring, Robert (Thesis advisor) / Hill, Gary (Committee member) / Rogers, Rodney (Committee member) / Schuring, Martin (Committee member) / Arizona State University (Publisher)
Created2016
ContributorsClements, Katrina (Performer) / ASU Library. Music Library (Publisher)
Created2018-03-15