Biochemistry, Biophysics, and Structural Biology Commons^™

Preparation Of A Flavonol Specific Glucosyltransferase Found In Grapefruit And Site-Directed Mutants For Protein Crystallization, Aaron Birchfield

Electronic Theses and Dissertations

This research was designed to determine the conditions necessary to remove c-myc and 6x-His tags from a flavonol specific glucosyltransferase found in grapefruit (CP3GT) using thrombin in preparation for crystallization. X-ray crystallography of CP3GT crystals may elucidate structural features that account for flavonol specificity in some glucosyltransferase enzymes. A thrombin cleavage site was inserted into WT CP3GT and one mutant. Recombinant CP3GT was expressed in yeast and purified. Optimal conditions for thrombin digestion were explored. Digestion with 100U of thrombin for 2 hours at 4^o C was optimal for removing tags from CP3GT. Storage at 4^o C for …

The Chelation Of Metal Ions By Vicibactin, A Siderophore Produced By Rhizobium Leguminosarum Atcc 14479, Joshua Stinnett

Undergraduate Honors Theses

Vicibactin is a small, high-affinity iron chelator produced by Rhizobium leguminosarum ATCC 14479. Previous work has shown that vicibactin is produced and secreted from the cell to sequester ferric iron from the environment during iron-deplete conditions. This ferric iron is then transported into the cell to be converted into ferrous iron. This study uses UV-Vis spectroscopy as well as ion trap-time of flight mass spectroscopy to determine that vicibactin does form a complex with copper(II) ions, however, at a much lower affinity than for iron(III). Stability tests have shown that the copper(II)-vicibactin complex is stable over time. The results of …

The Synthesis, Purification, And Characterization Of The P3 Peptoid, Myles Joyce

Chemical Engineering Undergraduate Honors Theses

The relatively young field of nano-systems has vast applications over various different fields. Peptoids (peptide analogues) and their uses are consistently investigated components of these nano-studies. A particularly interesting group of peptoids are able to self-assemble into secondary structures under the right conditions. The P3 peptoid is able to form microspheres if properly synthesized and purified. Microspheres are frequently studied due to their low surface area to volume ratios. This unique aspect of microspheres makes them an excellent candidate for drug delivery systems; however, these systems must also be robust so that the desired compound can be encapsulated within the …

Cloning, Expression, And Purification Of Fadk And Its Application In Coa-Rna Capture, Jon-Michael L. Stork

Honors Theses

Coenzyme A (CoA) is an important enzyme cofactor involved in acyl transfer reactions. Recently, CoA and its various thioesters were found to exist at the 5’ end of RNA molecules. Although the function of these molecules is still unknown due to difficulties in their isolation, their existence at the 5’ RNA end reveals potentially novel biological roles of RNA. We are exploiting the broad substrate tolerance of the medium chain fatty acid-coenzyme A ligase (FadK), hypothesizing that this enzyme will accept biotinylated medium chain fatty acid substrates, creating biotin-tagged CoA-RNA. This modified CoA-RNA can later be captured using streptavidin affinity …

The Role Of Parathyroid Hormone 1 Receptor (Pth1r) Signaling In Gastric Epithelial Homeostasis, Maram Maher Al-Hasan

Theses

In the stomach, the epithelial stem cells are responsible for glandular homeostasis. These stem cells continuously undergo cellular proliferation and differentiation to balance the death of senescent cells. Studies conducted on gastric cancers showed a significant increase in parathyroid hormone-like hormone (PTHLH). Moreover, both PTHLH along with parathyroid hormone (PTH) act as endogenous ligands for parathyroid hormone 1 receptor (PTH1R) which belongs to family B of G- protein-coupled receptors (GPCRs). This receptor functions by mediating many different signaling pathways and the most studied pathway is the stimulation of cyclic adenosine monophosphate (cAMP) as a second messenger.

There are very few …

Cellular Localization Of Rad51d Mutant Proteins And The Application Of Art To Increase Scientific Literacy In America, Claire L. Chabot

Senior Theses

Ovarian cancers are the leading cause of death from cancer of the female reproductive system. Approximately 50% of ovarian cancers have defects in the homologous recombination (HR) DNA repair pathway that is required for the repair of DNA double-stranded breaks. The status of HR genes, such as BRCA1, BRCA2, and the RAD51 family, contributes to ovarian cancer development as well as treatment decisions regarding chemotherapy, radiation, and immunotherapy. The overarching goal of this project is to identify new insights into HR that can integrate with Precision Medicine Initiatives and align with the goals of the Cancer Moonshot 2020 Program. I …

It’S Not Easy Being Green: A Study Of The Chemical Phenology Of The Eastern Hemlock Throughout A Growing Season, Nick Houseman, Evan Preisser

Senior Honors Projects

Phenology is the study of seasonally timed developmental events that are driven by environmental cues. Phenolic compounds and terpenoids are the two main classes of secondary metabolites present in conifer needles, and speaking generally, they correlate with needle age. Understanding the phenology of secondary metabolites is important, since variation in these compounds can affect a tree’s resistance to pest and pathogen attack. Terpenoids function in a wide array of ecological processes vital to conifer survival, including regulating forest dynamics through allelopathic inhibition of seed germination, altering rates of soil nutrient cycling and nitrification, and conferring resistance to pathogenic fungi and …

Targeting Sec61Α By Ipomoeassin F Leads To Highly Cytotoxic Effect, Zhijian Hu

Graduate Theses and Dissertations

Ipomoeassin F is a flagship congener of a resin glycoside family that inhibits growth of many tumor cell lines with only single-digital nanomolar IC50 values. However, biological and pharmacological mechanisms of ipomoeassin F have been undefined. To facilitate exploration of the biological and pharmacological properties, we performed sophisticate SAR (Structure–activity relationship) studies of ipomoeassin F to understand its pharmacophore and structure properties so that we can design favorable probes for further biological investigation. By applying appropriate deviates that possess fluorescent groups and similar bio-activity, the target protein was found to be localized in endoplasmic reticulum (ER). Through biotin affinity pull …

The Effects Of Gibberellic Acid And Auxin Hormones On Heliotropism In Sunflowers, Brandon Bernardo, Hagop S. Atamian

Student Scholar Symposium Abstracts and Posters

Sunflowers are one of many different plant species that are able to track and face the sun in order to optimize the amount of sunlight they are exposed to. This process of orienting towards the sun is called Heliotropism. Sunflowers are able to effectively orient themselves towards the sun because the growth rate on the East and West side of the stem alternates depending on the time of day. At dawn, the East facing stem will grow at a faster rate than the West facing side, resulting in the flower orienting towards the West. This alternating and uneven growth is …

Multiple Local Σ-Aromaticity Of Nonagermanide Clusters, Nikolay V. Tkachenko, Alexander I. Boldyrev

Chemistry and Biochemistry Faculty Publications

Nonagermanide clusters are widely used in inorganic synthesis and are actively studied by experimentalists and theoreticians. However, chemical bonding of such versatile species is still not completely understood. In our work we deciphered a bonding pattern for various experimentally obtained nonagermanide species. We localized the electron density via the AdNDP algorithm for the model structures ([Ge₉]⁴⁻, [Ge₉{P(NH₂)₂}₃]⁻, Cu[Ge₉{P(NH₂)₂}₃] and Cu(NHC)[Ge₉{P(NH₂)₂}₃]) and obtained a simple and chemically intuitive bonding pattern which …

Generalizing Metallocene Mechanochemistry To Ruthenocene Mechanophores, Ye Sha, Yudi Zhang, Enhua Xu, C Wayne Mcalister, Tianyu Zhu, Stephen L. Craig, Chuanbing Tang

Faculty Publications

Recent reports have shown that ferrocene displays an unexpected combination of force-free stability and mechanochemical activity, as it acts as the preferred site of chain scission along the backbone of highly extended polymer chains. This observation raises the tantalizing question as to whether similar mechanochemical activity might be present in other metallocenes, and, if so, what features of metallocenes dictate their relative ability to act as mechanophores. In this work, we elucidate polymerization methodologies towards main-chain ruthenocene-based polymers and explore the mechanochemistry of ruthenocene. We find that ruthenocene, in analogy to ferrocene, acts as a highly selective site of main …

Potential Interactions Between Glucocorticoid Receptors And L-Type Cav1.3 Ion Channels Expressed In Chinese Hamster Ovary (Cho) Cells, Mallory Soska

Undergraduate Distinction Papers

L-type calcium channels couple membrane depolarization to muscle contraction and aid in signal conduction between cells. Cav1.3 calcium channel isoform is expressed in neurons and endocrine cells. Nervous and endocrine tissues also express both Glucocorticoid (GR) and Mineralocorticoid receptors (MR) that are responsible for binding serum cortisol to induce immediate and long term changes within the cell. While neurons co-express both GRs and MRs, GR activation occurs in response to high concentrations of cortisol. There is a direct correlation between GR activation and the amplitude of calcium currents (Champeau, 2007). To determine potential interactions between the GR and the calcium …

Characterization Of The Perilipin 5 C-Terminus, Tim D. Clinton

Undergraduate Honors Thesis Projects

The crystallization of protein with subsequent X-Ray crystallography analysis is a powerful technique for uncovering a protein’s structure. This methodology is cutting edge and innovations are being made daily on how to best use the fundamentals of this technique for clearer characterizations. The protein Perilipin 5 could benefit from this technique as properties involving the structure of the molecule are still largely unknown. Knowing the structural aspects of perilipin 5 is important as several research efforts indicate that is a key factor in the regulation of lipolysis, or the breakdown of fats within a biological system. In the life sciences, …

The Effects Of Ultraviolet Light Exposure On The Activity Of Catalase In The Coelomocytes Of Sea Urchins Lytechinus Variegatus And Arbacia Punctulata, Kandis Arlinghaus

Undergraduate Theses

Many sea urchins play important ecological roles in their environments, and it is important to study the impacts of environmental stressors on their physiology. Ultraviolet radiation (UVR) exposure has significant negative impacts on marine organisms including an increase in reactive oxygen species (ROS). Oxidative damage by ROS at the cellular level can cause lipid peroxidation, DNA fragmentation, and even cell death that may result in inflammation or disease. To prevent this cellular damage, organisms generate enzymes, such as catalase, that breakdown ROS into harmless substances. Elevated catalase activities under UVB, a range of UVR from 280–315 nm, exposure have been …

Pbrm1 Regulates Stress Response In Epithelial Cells, Elizabeth G. Porter, Alisha Dhiman, Basudev Chowdhury, Benjamin C. Carter, Hang Lin, Jane C. Stewart, Majid Kazemian, Michael K. Wendt, Emily C. Dykhuizen

Department of Biochemistry Faculty Publications

Polybromo1 (PBRM1) is a chromatin remodeler subunit highly mutated in cancer, particularly clear cell renal carcinoma. PBRM1 is a member of the SWI/SNF subcomplex, PBAF (PBRM1-Brg1/Brm-associated factors), and is characterized by six tandem bromodomains. Here we establish a role for PBRM1 in epithelial cell maintenance through the expression of genes involved in cell adhesion, metabolism, stress response, and apoptosis. In support of a general role for PBRM1 in stress response and apoptosis, we observe that loss of PBRM1 results in an increase in reactive oxygen species generation and a decrease in cellular viability under stress conditions. We find that loss …

Interactions Of (My) 6 (M = Zn, Cd; Y = O, S, Se) Quantum Dots With N-Bases, Mariusz Michalczyk, Wiktor Zierkiewicz, Steve Scheiner

Chemistry and Biochemistry Faculty Publications

(MY)₆ clusters, with M = Zn and Cd and Y = O, S, Se, form double-layer drum-like structures containing M–Y covalent bonds. The positive regions near the M atoms attract the N atom of both NH₃ and NMe₃ so as to form a noncovalent M···N bond. This bond is quite strong, with interaction energies exceeding 35 kcal/mol. The bond strength diminishes with reduced electronegativity of the Y atom (O > S > Se) and is stronger for M = Zn than for Cd. Trimethylation of the base enhances the bond strength. The interaction is dominated by the electrostatic component …

Yield Stress Analysis With Dv-Iii Rheometer, Leah Margaret Cook

Honors Theses

The purpose of this thesis is to understand the behavior of everyday substances (toothpaste, lotions, ect.) and chemicals through the analysis of yield stress. By measuring the flow properties of these products, one can determine the stability, uses, and other properties of a substance. Yield stress is experienced everyday from squeezing a tube of toothpaste, to stirring a batch of cake batter, or canning products in a factory. Yield stress along with viscosity helps us predict the behavior of chemicals, cosmetics, and food that we handle daily. Yield stress, torque, and strain values were measured with a DV-III Rheometer and …

Gamete Nuclear Migration In Animals And Plants, Umma Fatema, Mohammad F. Ali, Zheng Hu, Anthony J. Clark, Tomokazu Kawashima

Plant and Soil Sciences Faculty Publications

The migration of male and female gamete nuclei to each other in the fertilized egg is a prerequisite for the blending of genetic materials and the initiation of the next generation. Interestingly, many differences have been found in the mechanism of gamete nuclear movement among animals and plants. Female to male gamete nuclear movement in animals and brown algae relies on microtubules. By contrast, in flowering plants, the male gamete nucleus is carried to the female gamete nucleus by the filamentous actin cytoskeleton. As techniques have developed from light, electron, fluorescence, immunofluorescence, and confocal microscopy to live-cell time-lapse imaging using …

Department Of Chemistry And Biochemistry 24th Departmental Symposium In Conjunction With The Petersheim Academic Exposition, Seton Hall University

Petersheim Academic Exposition

No abstract provided.

Semisynthetic Aurones Inhibit Tubulin Polymerization At The Colchicine-Binding Site And Repress Pc-3 Tumor Xenografts In Nude Mice And Myc-Induced T-All In Zebrafish, Yanqi Xie, Liliia M. Kril, Tianxin Yu, Wen Zhang, Mykhaylo S. Frasinyuk, Svitlana P. Bondarenko, Kostyantyn M. Kondratyuk, Elizabeth Hausman, Zachary M. Martin, Przemyslaw P. Wyrebek, Xifu Liu, Agripina G. Deaciuc, Linda P. Dwoskin, Jing Chen, Haining Zhu, Chang-Guo Zhan, Vitaliy M. Sviripa, Jessica S. Blackburn, David S. Watt, Chunming Liu

Molecular and Cellular Biochemistry Faculty Publications

Structure-activity relationships (SAR) in the aurone pharmacophore identified heterocyclic variants of the (Z)-2-benzylidene-6-hydroxybenzofuran-3(2H)-one scaffold that possessed low nanomolar in vitro potency in cell proliferation assays using various cancer cell lines, in vivo potency in prostate cancer PC-3 xenograft and zebrafish models, selectivity for the colchicine-binding site on tubulin, and absence of appreciable toxicity. Among the leading, biologically active analogs were (Z)-2-((2-((1-ethyl-5-methoxy-1H-indol-3-yl)methylene)-3-oxo-2,3-dihydrobenzofuran-6-yl)oxy)acetonitrile (5a) and (Z)-6-((2,6-dichlorobenzyl)oxy)-2-(pyridin-4-ylmethylene)benzofuran-3(2H)-one (5b) that inhibited in vitro PC-3 prostate cancer cell proliferation with IC₅₀ values below 100 nM. A xenograft study in nude mice using …