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Proteins continually and naturally incur evolutionary selection through mutagenesis that optimizes their fitness, which is primarily determined by their function. It is known that allosteric regulation alters a protein's conformational dynamics leading to functional changes. We have computationally introduced a mutation at a predicted regulatory site of a short, 46

Proteins continually and naturally incur evolutionary selection through mutagenesis that optimizes their fitness, which is primarily determined by their function. It is known that allosteric regulation alters a protein's conformational dynamics leading to functional changes. We have computationally introduced a mutation at a predicted regulatory site of a short, 46 residue-long, protein interaction module composed of a WW domain and corresponding polyproline ligand (PDB id: 1k9r). The dynamic flexibility index (DFI) was computed for the binding site of the wild type and mutant WW domains to quantify the mutations effect on the rigidity of the binding pocket. DFI is used as a metric to quantify the resilience of a given position to perturbation along the chain. Using steered molecular dynamics (SMD), we also measure the effect of the point mutation on allosteric regulation by approximating the binding free energy of the system calculated using Jarzynski's Equality. Calculation of the DFI shows that the overall flexibility of the protein complex increases as a result of the distal point mutation. Total change in DFI percentile of the binding site showed a 0.067 increase suggesting an allosteric, loss of function mutation. Furthermore, we see that the change in the binding free energy is greater for that of the mutated complex supporting the idea that an increase in flexibility is correlated to a decrease in proteinlig and binding affinity. We show that sequence mutation of an allosteric site affects the mechanical stability and functionality of the binding pocket.
ContributorsMarianchuk, Tegan (Author) / Ozkan, Sefika (Thesis director) / Ros, Robert (Committee member) / Barrett, The Honors College (Contributor) / Department of Physics (Contributor)
Created2018-05
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The purpose of this paper is to provide an analysis of entanglement and the particular problems it poses for some physicists. In addition to looking at the history of entanglement and non-locality, this paper will use the Bell Test as a means for demonstrating how entanglement works, which measures the

The purpose of this paper is to provide an analysis of entanglement and the particular problems it poses for some physicists. In addition to looking at the history of entanglement and non-locality, this paper will use the Bell Test as a means for demonstrating how entanglement works, which measures the behavior of electrons whose combined internal angular momentum is zero. This paper will go over Dr. Bell's famous inequality, which shows why the process of entanglement cannot be explained by traditional means of local processes. Entanglement will be viewed initially through the Copenhagen Interpretation, but this paper will also look at two particular models of quantum mechanics, de-Broglie Bohm theory and Everett's Many-Worlds Interpretation, and observe how they explain the behavior of spin and entangled particles compared to the Copenhagen Interpretation.

ContributorsWood, Keaten Lawrence (Author) / Foy, Joseph (Thesis director) / Hines, Taylor (Committee member) / Department of Physics (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Description

Lyme disease is a common tick-borne illness caused by the Gram-negative bacterium Borrelia burgdorferi. An outer membrane protein of Borrelia burgdorferi, P66, has been suggested as a possible target for Lyme disease treatments. However, a lack of structural information available for P66 has hindered attempts to design medications to target

Lyme disease is a common tick-borne illness caused by the Gram-negative bacterium Borrelia burgdorferi. An outer membrane protein of Borrelia burgdorferi, P66, has been suggested as a possible target for Lyme disease treatments. However, a lack of structural information available for P66 has hindered attempts to design medications to target the protein. Therefore, this study attempted to find methods for expressing and purifying P66 in quantities that can be used for structural studies. It was found that by using the PelB signal sequence, His-tagged P66 could be directed to the outer membrane of Escherichia coli, as confirmed by an anti-His Western blot. Further attempts to optimize P66 expression in the outer membrane were made, pending verification via Western blotting. The ability to direct P66 to the outer membrane using the PelB signal sequence is a promising first step in determining the overall structure of P66, but further work is needed before P66 is ready for large-scale purification for structural studies.

ContributorsRamirez, Christopher Nicholas (Author) / Fromme, Petra (Thesis director) / Hansen, Debra (Committee member) / Department of Physics (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
Description

The self-assembly of strongly-coupled nanocrystal superlattices, as a convenient bottom-up synthesis technique featuring a wide parameter space, is at the forefront of next-generation material design. To realize the full potential of such tunable, functional materials, a more complete understanding of the self-assembly process and the artificial crystals it produces is

The self-assembly of strongly-coupled nanocrystal superlattices, as a convenient bottom-up synthesis technique featuring a wide parameter space, is at the forefront of next-generation material design. To realize the full potential of such tunable, functional materials, a more complete understanding of the self-assembly process and the artificial crystals it produces is required. In this work, we discuss the results of a hard coherent X-ray scattering experiment at the Linac Coherent Light Source, observing superlattices long after their initial nucleation. The resulting scattering intensity correlation functions have dispersion suggestive of a disordered crystalline structure and indicate the occurrence of rapid, strain-relieving events therein. We also present real space reconstructions of individual superlattices obtained via coherent diffractive imaging. Through this analysis we thus obtain high-resolution structural and dynamical information of self-assembled superlattices in their native liquid environment.

ContributorsHurley, Matthew (Author) / Teitelbaum, Samuel (Thesis director) / Ginsberg, Naomi (Committee member) / Kirian, Richard (Committee member) / Barrett, The Honors College (Contributor) / Department of Physics (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Historical, Philosophical & Religious Studies, Sch (Contributor)
Created2023-05
Description

This qualitative study sought to investigate the potential reaction between the 3,3',5,5'-tetramethylbenzidine (TMB) radical and LAF-1 RGG, the N-terminus domain of an RNA helicase which functions as a coacervating intrinsically disordered protein. The study was performed by adding horseradish peroxidase to a solution containing TMB and either LAF-1 or tyrosine

This qualitative study sought to investigate the potential reaction between the 3,3',5,5'-tetramethylbenzidine (TMB) radical and LAF-1 RGG, the N-terminus domain of an RNA helicase which functions as a coacervating intrinsically disordered protein. The study was performed by adding horseradish peroxidase to a solution containing TMB and either LAF-1 or tyrosine in various concentrations, and monitoring the output through UV-Vis spectroscopy. The reacted species was also analyzed via MALDI-TOF mass spectrometry. UV-Vis spectroscopic monitoring showed that in the presence of LAF-1 or tyrosine, the reaction between HRP and TMB occurred more quickly than the control, as well as in the highest concentration of LAF-1, the evolution of a peak at 482 nm. The analysis through MALDI-TOF spectrometry showed the development of a second peak likely due to the reaction between LAF-1 and TMB, as the Δ between the peaks is 229 Da and the size of the TMB species is 240 Da.

ContributorsDavis, Morgan (Author) / Ghirlanda, Giovanna (Thesis director) / Heyden, Matthias (Committee member) / Mazor, Yuval (Committee member) / Barrett, The Honors College (Contributor) / Department of Physics (Contributor) / School of Molecular Sciences (Contributor)
Created2022-12
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Description
Carbon allotropes are the basis for many exciting advancements in technology. While sp² and sp³ hybridizations are well understood, the sp¹ hybridized carbon has been elusive. However, with recent advances made using a pulsed laser ablation in liquid technique, sp¹ hybridized carbon allotropes have been created. The fabricated carbon chain

Carbon allotropes are the basis for many exciting advancements in technology. While sp² and sp³ hybridizations are well understood, the sp¹ hybridized carbon has been elusive. However, with recent advances made using a pulsed laser ablation in liquid technique, sp¹ hybridized carbon allotropes have been created. The fabricated carbon chain is composed of sp¹ and sp³ hybridized bonds, but it also incorporates nanoparticles such as gold or possibly silver to stabilize the chain. The polyyne generated in this process is called pseudocarbyne due to its striking resemblance to the theoretical carbyne. The formation of these carbon chains is yet to be fully understood, but significant progress has been made in determining the temperature of the plasma in which the pseudocarbyne is formed. When a 532 nm pulsed laser with a pulsed energy of 250 mJ and pulse length of 10ns is used to ablate a gold target, a peak temperature of 13400 K is measured. When measured using Laser-Induced Breakdown spectroscopy (LIBS) the average temperature of the neutral carbon plasma over one second was 4590±172 K. This temperature strongly suggests that the current theoretical model used to describe the temperature at which pseudocarbyne generates is accurate.
ContributorsWala, Ryland Gerald (Co-author) / Wala, Ryland (Co-author) / Sayres, Scott (Thesis director) / Steimle, Timothy (Committee member) / Drucker, Jeffery (Committee member) / Historical, Philosophical & Religious Studies (Contributor) / Dean, W.P. Carey School of Business (Contributor) / Department of Physics (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2019-05