Biochemistry, Biophysics, and Structural Biology Commons^™

Characterization Of The Product Specificity And Kinetic Mechanism Of Protein Arginine Methyltransferase 1, Shanying Gui

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Protein enzymes perform a vast array of functions within living organisms, catalyzing various metabolic reactions including DNA replication, DNA repair, protein synthesis, etc. In order to maintain proper cellular functions, enzymes need to be accurately regulated under different circumstances. Specifically, enzymes can be modified after their creation to give them additional functions. These modifications can do a variety of things including activating (turning on) or inactivating (turning off) an enzyme, changing what proteins or molecules can interact with the enzyme, changing the enzyme’s location in the cell, and/or targeting the enzyme for destruction. This dissertation focuses on a single class …

Investigation Of The Oxidation/Reduction Of Prmt1, Substrate Interaction With Prmt1, And The Role Of Arginine Methylation In Rna Surveillance, Damon V. Nitzel

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Enzymes are the machines of our cells. Just like machines, it takes a lot of energy to create them, and they then serve only the function they were created for. If we want to change the function of a machine, we need to modify it. Similarly, enzymes can be modified after their creation to give them additional function. These modifications can do a variety of things including activating (on) or inactivating (off) an enzyme, changing the enzyme’s location in the cell, and targeting the enzyme for destruction. This thesis focuses on a single class of enzymes, protein arginine methyltransferases (PRMTs), …

Mesobiliverdin Ixα Enhances Rat Pancreatic Islet Yield And Function, Taihei Ito, Dong Chen, Cheng-Wei Tom Chang, Takashi Kenmochi, Tomonori Saito, Satoshi Suzuki, Jon Y. Takemoto

Chemistry and Biochemistry Faculty Publications

The aims of this study were to produce mesobiliverdin IXα, an analog of anti-inflammatory biliverdin IXα, and to test its ability to enhance rat pancreatic islet yield for allograft transplantation into diabetic recipients. Mesobiliverdin IXα was synthesized from phycocyanobilin derived from cyanobacteria, and its identity and purity were analyzed by chromatographic and spectroscopic methods. Mesobiliverdin IXα was a substrate for human NADPH biliverdin reductase. Excised Lewis rat pancreata infused with mesobiliverdin IXα and biliverdin IXα-HCl (1–100 μM) yielded islet equivalents as high as 86.7 and 36.5%, respectively, above those from non-treated controls, and the islets showed a high degree of …

The Role Of The Global Cryosphere On The Fate Of Organic Contaminants., Amanda M. Grannas, Christian Bogdal, Kimberly J. Hageman, Frank Wania

Chemistry and Biochemistry Faculty Publications

The cryosphere is an important component of global organic contaminant cycles. Snow is an efficient scavenger of atmospheric organic pollutants while a seasonal snowpack, sea ice, glaciers and ice caps are contaminant reservoirs on time scales ranging from days to millennia. Important physical and chemical processes occurring in the various cryospheric compartments impact contaminant cycling and fate. A variety of interactions and feedbacks also occur within the cryospheric system, most of which are susceptible to perturbations due to climate change. In this article, we review the current state of knowledge regarding the transport and processing of organic contaminants in the …

Characterization Of A Novel Fusion Protein Of Ipab And Ipad Of Shigella And Its Potential As A Pan-Shigella Vaccine, F. J. Martinez-Becerra, X. Chen, Nicholas E. Dickenson, S. P. Choudhari, K. Harrison, J. D. Clements, W. D. Picking, L. L. Van De Verg, R. I. Walker, W. L. Picking

Chemistry and Biochemistry Faculty Publications

Shigellosis is an important disease in the developing world, where about 90 million people become infected with Shigella spp. each year. We previously demonstrated that the type three secretion apparatus (T3SA) proteins IpaB and IpaD are protective antigens in the mouse lethal pulmonary model. In order to simplify vaccine formulation and process development, we have evaluated a vaccine design that incorporates both of these previously tested Shigella antigens into a single polypeptide chain. To determine if this fusion protein (DB fusion) retains the antigenic and protective capacities of IpaB and IpaD, we immunized mice with the DB fusion and compared …

The Magnitude And Mechanism Of Charge Enhancement Of Ch∙∙O H-Bonds, U. Adhikari, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

Quantum calculations find that neutral methylamines and thioethers form complexes, with N-methylacetamide (NMA) as proton acceptor, with binding energies of 2–5 kcal/mol. This interaction is magnified by a factor of 4–9, bringing the binding energy up to as much as 20 kcal/mol, when a CH3+ group is added to the proton donor. Complexes prefer trifurcated arrangements, wherein three separate methyl groups donate a proton to the O acceptor. Binding energies lessen when the systems are immersed in solvents of increasing polarity, but the ionic complexes retain their favored status even in water. The binding energy is reduced when the methyl …

Caehnorhabditis Elegans: A Low-Cost In Vivo Animal Model For Efficacy Studies Of Novel Antibiotics, Rylee Ann Gregory

Undergraduate Honors Capstone Projects

Since the 1940s, antibiotics have greatly reduced the adverse effects of infectious diseases caused by microbes. However, due to excessive, and often incorrect, use of known antibiotics, many organisms have adapted antibiotic resistance. Currently, over 70% of known infectious bacteria are resistant to at least one antibiotic. In the U.S. , 90,000 deaths occur each year due to infection by bacteria resistant to antibiotics. This number has increased by nearly 75,000 in the last 20 years. It is necessary, therefore, to continue developing new antibiotics in an effort to keep up with increasing antibiotic resistance. Traditional in vitro and whole …

Locking Down The N-Terminus Of Prmt1 In Order To Assess The Role Of Motion In Activity, Taylor James Rasmussen

Undergraduate Honors Capstone Projects

Protein arginine methyltransferases (PRMTs) are involved In many major biological pathways in the human body. Processes that demonstrate arginine methylation by PRMTs include, but are not limited to, histone modification, DNA transcription, and post-translational protein modifications. Although recent research has allowed the identification of several PRMT isoforms and exposed their involvement in these processes, relatively little is known about the details of how these enzymes perform their biochemical duties. It is currently hypothesized that the N-terminus of PRMT variant I is involved in recognizing substrates and aiding in catalysis by virtue of a change in its conformation. To understand how …

Biochemical Characterization Of Sac9, A Putative Phosphoinositide Phosphatase In Arabidopsis Thaliana, And Its Role In Cellular Abnormalities, Almut H. Vollmer

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Since the early colonization of land by fungi and plants some 700 million years ago, plants have been continuously faced with changes in their environment. Unlike animals, plants are not free to move about, and can therefore not evade many stress factors. How plants sense and respond to their environment has been of interest not only to scientific research but also in more practical applications such as agriculture.

Signals (such as light or salinity) from the outside of plant cells trigger a flow of information to the inside of the cell. The final target for most of the information is …

1. Improving The Yield Of Biodiesel From Microalgae And Other Lipids. 2. Studies Of The Wax Ester Biosynthetic Pathway And Potential Biotechnological Application, Bradley D. Wahlen

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The production of biofuels and oleochemicals from renewable sources offers an opportunity to reduce our dependence on fossil fuels. The work contained in this dissertation has focused on developing and improving methods for the production of biodiesel from non-traditional feedstocks and understanding biosynthetic pathways that result in the production of oleochemicals and fuels.

Pure vegetable oil can account for 70-80% of the total cost of biodiesel production. Many low-cost oils contain high amounts of free fatty acids, which are unsuitable for base-catalyzed transesterification. Herein an approach is described that efficiently accomplishes the simultaneous esterification and transesterification of both free fatty …

Structural And Mechanistic Investigations Of Phosphothreonine Lyase Class Of Enzymes, Alok Gopalkrishna Shenoy

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Phosphorylation and dephosphorylation are a highly pervasive mechanism in biology that is used by the cell to modulate enzymes and proteins. The presence of a phosphate group can activate or deactivate an enzyme. The phosphate group is linked to a protein by a phosphoester bond that is known to be highly stable in cytoplasmic pH range. Thus the breaking and formation of these bonds need to be effected by enzymes.

Recent discovery of the activity carried out by certain virulence related proteins (OspF released by Shigella and SpvC released by Salmonella) have resulted in a necessity to create a new …

Detailed Comparison Of The Pnicogen Bond With Chalcogen, Halogen And Hydrogen Bonds, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

The characteristics of the pnicogen bond are explored using a variety of quantum chemical techniques. In particular, this interaction is compared with its halogen and chalcogen bond cousins, as well as with the more common H-bond. In general, these bonds are all of comparable strength. More specifically, they are strengthened by the presence of an electronegative substituent on the electron-acceptor atom, and each gains strength as one moves down the appropriate column of the periodic table, for example, from N to P to As. These noncovalent bonds owe their stability to a mixture in nearly equal parts of electrostatic attraction …

First Steps In Growth Of A Polypeptide Toward Β-Sheet Structure, U. Adhikari, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

The full conformational energy surface is examined for a molecule in which a dipeptide is attached to the same spacer group as another peptide chain, so as to model the seminal steps of β-sheet formation. This surface is compared with the geometrical preferences of the isolated dipeptide to extract the perturbations induced by interactions with the second peptide strand. These interpeptide interactions remove any tendency of the dipeptide to form a C5 ring structure, one of its two normally stable geometries. A C7 structure, the preferred conformation of the isolated dipeptide, remains as the global minimum in the full molecule. …

Extrapolation To The Complete Basis Set Limit For Binding Energies Of Noncovalent Interactions, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

A means of extrapolating from double and triple-valence basis sets to a complete basis set is examined in the context of the pnicogen bonds in the BH2P⋯NH3 complexes, with B = CH3, H, NH2, CF3, OH, Cl, F, and NO2. Binding energies converge smoothly, and the trends for the various substituents B are unaffected by the basis set size, extrapolation, or level of inclusion of electron correlation, including MP2 and CCSD(T). The approach appears to be successful also for H-bonded systems, in particular the water dimer. In the event that full extrapolation within the context of CCSD(T) is not feasible, …

Sensitivity Of Noncovalent Bonds To Intermolecular Separation: Hydrogen, Halogen, Chalcogen, And Pnicogen Bonds, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

It is well known that noncovalent bonds are weakened when stretched from their equilibrium intermolecular separation. Quantum chemical calculations are used to examine and compare the sensitivity to stretches of hydrogen, halogen, chalcogen, and pnicogen bonds. NH3 was taken as the universal electron donor, paired with HOH and FH in H-bonds, as well as with FPH2, FSH, and FCl. Even though the binding energies span a wide range, stretching the intermolecular separation by 1 Å cuts this quantity by the same proportion, roughly in half, for each system. Taking the sum of van der Waals radii as an arbitrary cutoff, …

Preferred Configurations Of Peptide-Peptide Interactions, U. Adhikari, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

The natural and fundamental proclivities of interaction between a pair of peptide units are examined using high-level ab initio calculations. The NH···O H-bonded structure is found to be the most stable configuration of the N-methylacetamide (NMA) model dimer, but only slightly more so than a stacked arrangement. The H-bonded geometry is destabilized by only a small amount if the NH group is lifted out of the plane of the proton-accepting amide. This out-of-plane motion is facilitated by a stabilizing charge transfer from the CO π bond to the NH σ* antibonding orbital. The parallel and antiparallel stacked dimers are nearly …

Evaluation Of Dft Methods To Study Reactions Of Benzene With Oh Radical, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

Several density functional theory” (DFT) methods are applied to two different reaction channels involving OH• + C6H6, and the results compared with high-level ab initio calculations. The OH• adds directly to one C atom in the first channel, first forming an encounter complex with the OH• poised above the aromatic plane. B3LYP, BH&HLYP, and MPW1K compute an accurate estimate of the overall exothermicity, whereas M05-2X, PBE0, and PBEPBE overestimate this quantity to some degree. With the exceptions of PBEPBE and PBE0, the other methods produce an acceptable barrier to addition. All approaches except BH&HLYP correctly predict an exothermic H• abstraction, …

Contributions Of Various Noncovalent Bonds To The Interaction Between An Amide And S-Containing Molecules, U. Adhikari, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

N-Methylacetamide, a model of the peptide unit in proteins, is allowed to interact with CH3SH, CH3SCH3, and CH3SSCH3 as models of S-containing amino acid residues. All of the minima are located on the ab initio potential energy surface of each heterodimer. Analysis of the forces holding each complex together identifies a variety of different attractive forces, including SH⋅⋅⋅O, NH⋅⋅⋅S, CH⋅⋅⋅O, CH⋅⋅⋅S, SH⋅⋅⋅π, and CH⋅⋅⋅π H-bonds. Other contributing noncovalent bonds involve charge transfer into σ* and π* antibonds. Whereas some of the H-bonds are strong enough that they represent the sole attractive force in several dimers, albeit not usually in the …

Effects Of Carbon Chain Substituent On The P∙∙∙N Noncovalent Bond, U. Adhikari, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

The effects of carbon chains placed on the electron-accepting P atom of a P⋯N bond are examined via ab initio calculations. Saturated alkyl groups have a mild weakening effect, regardless of chain length. In contrast, incorporation of double bonds into the chain strengthens the interaction, Ctriple bond; length of mdashC triple bonds even more so. These effects are only slightly enhanced by additional conjugated double bonds or an aromatic ring. Placing F atoms onto the carbon chains strengthens the P⋯N bond, but only by a small amount, which wanes as the F atom is displaced further from the P along …

Interfamily Variation In Amphibian Early Life-History Traits: Raw Material For Natural Selection?, Gareth R. Hopkins, Brian G. Gall, Susannah S. French, Edmund D. Brodie Jr.

Biology Faculty Publications

The embryonic development and time to hatching of eggs can be highly adaptive in some species, and thus under selective pressure. In this study, we examined the underlying interfamily variation in hatching timing and embryonic development in a population of an oviparous amphibian, the rough-skinned newt (Taricha granulosa). We found significant, high variability in degree of embryonic development and hatching timing among eggs from different females. Patterns of variation were present regardless of temperature.We also could not explain the differences among families by morphological traits of the females or their eggs. This study suggests that the variation necessary for natural …

Studies Of The Distinguishing Features Of Nadph:2-Ketopropyl-Coenzyme M Oxidoreductase/Carboxylase, An Atypical Member Of The Disulfide/ Oxidoreductase Family Of Enzymes, Melissa A. Beighley-Kofoed

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The metabolism of propylene in Xanthobacter autotrophicus occurs via epoxypropane formation and subsequent metabolism by a three-step, four-enzyme pathway, utilizing the atypical cofactor Coenzyme M (CoM) to form acetoacetate. The last step in the epoxide carboxylase pathway is catalyzed by a distinctive member of the disulfide oxidoreductase (DSOR) family of enzymes, NADPH:2-ketopropyl CoM oxidoreductase/carboxylase (2-KPCC). 2-KPCC catalyzes the unorthodox cleavage of a thioether bond and successive carboxylation of the substrate. The focus of the research presented in this dissertation aims to elucidate the details of 2-KPCC that allow it to perform chemistry unconventional for typical DSOR members. Sitedirected mutagenesis was …

A New Noncovalent Force: Comparison Of P∙∙∙N Interaction With Hydrogen And Halogen Bonds, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

When PH(3) is paired with NH(3), the two molecules are oriented such that the P and N atoms face one another directly, without the intermediacy of a H atom. Quantum calculations indicate that this attraction is due in part to the transfer of electron density from the lone pair of the N atom to the σ(∗) antibond of a P-H covalent bond. Unlike a H-bond, the pertinent hydrogen is oriented about 180° away from, instead of toward, the N, and the N lone pair overlaps with the lobe of the P-H σ(∗) orbital that is closest to the P. In …

Kinetic, Mechanistic, And Structural Investigation Of Features Controlling Stereoselectivity Of (R)- And (S)-Hydroxypropyl Com Dehydrogenases From Xanthobacter Autrophicus Strain Py2, Dariusz Adam Sliwa

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Enantiopure alcohols are valuable intermediates in fine organic synthesis, in particular for preparation of biologically active compounds. The necessity of preparing single enantiomer drugs in an optically pure form has triggered much research, especially in the pharmaceutical industry. The biocatalytical production of chiral alcohols by alcohol dehydrogenase enzymes is characterized by the asymmetric reduction of the corresponding ketones, usually with high degree of stereoselectivity. The commercial value of the enzymes as stereoselective biocatalysts has been a significant driving force in understanding features that control their mechanism of catalysis and stereoselectivity. This work focuses on two enantiocomplementary dehydrogenase enzymes ((R …

Properties Of Microbes In Natural Fire Burn Soils, Holly Anderson

Undergraduate Honors Capstone Projects

Soil underlying a natural fire develops a hydrophobic soil sub-layer. This hydrophobicity decreases with time although the mechanisms are unresolved, but are thought to be biotic and abiotic. Some of the compounds accounting for hydrophobicity are polycyclic aromatic hydrocarbons (PAHs) of which pyrene is an example.

Bacteria that grew on pyrene were isolated from burned soils at two sites in Utah in order to analyze the biotic microbial degradation of the hydrophobic soil sub-layer. The two sites were Wood Camp (Logan, UT) and Milford Flats (Central UT). Identifications of the genera of nine isolated bacteria were made through l6S rRNA …

Host Signaling Response To Adhesion Of Bifidobacterium Infantis, Reed N. Gann

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Investigations of the molecular binding partners of the probiotic bacterium Bifidobacterium longum subspecies infantis (B. infantis) and the pathogen Salmonella enterica subspecies enterica serovar Typhimurium LT2 (Salmonella ser. Typhimurium) found that these two very different bacteria bind gangliosides. However, these organisms lead to completely different host health outcomes when present in the gut. B. infantis is the founding microbial population in the intestinal tract of breast-fed infants. S. typhimurium is the most important food-borne pathogen that results in humans. This study used an in vitro gut epithelial cell model to examine the host cellular response to adhesion …

Carboxylases Involved In Microbial Acetone And Acetophenone Metabolism, Ameya Mashruwala

All Graduate Plan B and other Reports, Spring 1920 to Spring 2023

A number of bacteria are capable of growth with acetone and acetophenone as their sole sources of carbon and/or energy. The pathways and enzymes involved in the transformation of these molecules into useable carbon and energy are unique. Among these are two novel enzymes, acetone carboxylase and acetophenone carboxylase, which represent a fundamentally novel classes of carboxylases.

The initial step in acetone metabolism, in X. autotrophicus st Py2, R. capsulatus st B10 and R. rhodochrous, is the thermodynamically unfavorable reaction to yield acetoacetate. This step is catalyzed by the enzyme acetone carboxylase and is coupled with the unprecedented, concomitant hydrolysis …

Genetic And Biochemical Studies Of Plasmid Pir52-1 In Lactobacillus Helveticus, Cody Alexander Tramp

Undergraduate Honors Capstone Projects

Lactobacillus helveticus is a species of lactic acid bacteria. These bacteria, which produce lactic acid as a major product of carbohydrate metabolism, are used industrially to produce cheese and other fermented dairy products (Ebringer et al., 2008). Many species of lactic acid bacteria also possess probiotic characteristics and when ingested potentially confer increased immune function, regulate gut microbiota, and improve digestion in the host (Reid, 2008). Genetic studies of these probiotic effects and other characteristics of L. helveticus and related species have been hindered by the lack of stably replicating plasmid vectors. Plasmid vectors for several Lactobacillus species have been …

Physiological And Molecular Function Of Hap3b In Flowering Time Regulation And Cold Stress Response, Mingxiang Liang

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Heme-activated proteins (HAPs) are transcription factors that have multiple roles in plant growth and development, such as embryogenesis, flowering time control, and drought tolerance.

In the present study I found that HAP3b was also involved in controlling response to cold stress. Transcript profiling and gene expression analyses indicated that HAP3b repressed the CBF3 regulon under normal growth conditions. As a result, plants with HAP3b-overexpressed showed decreased survival rates while plants homozygous for the null allele hap3b showed an improved freezing tolerance compared to wild-type plants.

To understand the mechanism of HAP3b in Arabidopsis, i.e. whether it also acts …

Transesterification And Recovery Of Intracellular Lipids Using A Single Step Reactive Extraction, Daniel R. Nelson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A single-step, extractive reaction for extraction of lipids such as biodiesel components, omega-3 fatty acids, or other triglycerides from microbial cells was examined. Conventional methods for lipid extraction use toxic solvents, and require multiple steps and long processing times. When the goal is to produce fatty acid methyl esters or FAMEs, the extracted lipids are subjected to a separate transesterification reaction with simple alcohols in the presence of an acid or base catalyst. A simplified, single-step reactive extraction method can be applied that combines the sequential extraction followed by transesterification using acidified alcohols - a process known as in situ …

A Synthetic Biological Engineering Approach To Secretion- Based Recovery Of Polyhydroxyalkanoates And Other Cellular Products, Elisabeth Linton

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The costs associated with cellular product recovery commonly account for as much as 80% of the total production expense. As a specific example, significant recovery costs limit commercial use of polyhydroxyalkanoates (PHA), which comprise a class of microbially-accumulated polyesters. PHAs are biodegradable compounds that are of interest as a sustainable alternative to petrochemically-derived plastics. Secretion-based recovery of PHAs was studied to decrease PHA production costs. Type I and II secretory pathways are commonly used for the translocation of recombinant proteins out of the cytoplasm of E. coli. Proteins were targeted for translocation using four signal peptides (HlyA, TorA, GeneIII, …