Biochemistry, Biophysics, and Structural Biology Commons^™

Characterization Of The Catalytic Mechanism Of The Carboxyltransferase Component Of Escherichia Coli Acetyl-Coa Carboxylase Leading To Rational Inhibitor Identification And The Development Of Enzymatic Mimics, Nabil K. Thalji

Honors Capstones

No abstract provided.

Generation Of An Fsh Inducible Inh-[Alpha]-Flag-Gg-Icer I[Gamma] Ovarian Specific Gene Construct, Goce Bogdanoski

Theses, Dissertations and Culminating Projects

Avian animal models have led to many essential discoveries in molecular biology and biochemistry, including the discovery of vitamins, the chemistry behind vision and the development of transgenic hens as natural bioreactors for pharmaceutically relevant proteins. Hyperovulation in domesticated hens is considered a highly desirable genetic trait, a feature that has been explored in multiple studies and remains the focus of many biotech companies. Unlocking the molecular mechanism that controls ovulation may provide a basis for development in new reproductive technologies.

Previous research done on rodents provided data and identified a single transcription factor, the Inducible cAMP Early Repressor (ICER), …

Characterization Of The Substrate Specificity And Mechanism Of Protein Arginine Methyltransferase 1, Whitney Lyn Wooderchak

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Protein arginine methyltransferases (PRMTs) posttranslationally modify protein arginine residues. Type I PRMTs catalyze the formation of monomethylarginine (MMA) and asymmetric dimethylarginine (ADMA) via methyl group transfer from S-adenosyl methionine onto protein arginine residues. Type II PRMTs generate MMA and symmetric dimethylarginine. PRMT-methylation affects many biological processes. Although PRMTs are vital to normal development and function, PRMT-methylation is also linked to cardiovascular disease, stroke, multiple sclerosis, and cancer.

Thus far, nine human PRMT isoforms have been identified with orthologues present in yeast, plants, and fish. PRMT1 predominates, performing an estimated 85% of all protein arginine methylation in vivo. Yet, the substrate …

Molecular Coevolution Of Pacific Northwest Hantaviruses And Their Host, The Deer Mouse, Peromyscus Maniculatus, Philip Darren Jones

Dissertations and Theses

Sin Nombre virus (SNV, family Bunyaviridae, genus Hantavirus), hosted by the deer mouse, Peromyscus maniculatus (family Cricetidae, Subfamily Neotominae), is the primary etiological agent of Hantavirus pulmonary syndrome (HPS) in the western United States. HPS, with known pathogenicity only to humans and for which there is no cure or prophylaxis, affects the epithelium of the lungs by making the capillaries leaky, thereby resulting in bilateral infiltrates, and eventually leading to respiratory failure and death by drowning in approximately 38% of hospitalized patients.

In the Americas, Peromyscus has been co-evolving with hantaviruses for approximately 12–20 million years, since the first …

A Watershed Analysis Of Salmon Lake And Mcgrath Pond: Implications For Water Quality And Land Use Management, Colby Environmental Assessment Team, Colby College, Problems In Environmental Science Course (Biology 493), Colby College

Colby College Watershed Study: Salmon Lake and McGrath Pond (2009, 1993)

In the summer and fall of 2009, the Colby Environmental Assessment Team (CEAT) studied the water quality of Salmon Lake and McGrath Pond, located in the Belgrade Lakes Region of Maine. The physical, chemical and biological characteristics of water quality were measured and analyzed to evaluate the current health of these lakes. Water quality data collected during the summer and fall of 2009 were compared with data from previous years to study the historic water quality trends. Land use patterns in the Salmon/McGrath watershed were also examined to investigate their impact on the lake water quality. The water quality trends …

Structure-Function Analysis Of Endoproteolytic Cleavage: Site-Directed Mutagenesis Studies Of The Α-Msh Cleavage Site In Silurana Tropicalis Pomc, Quinn Kun Chen

Electronic Theses and Dissertations

The maturation process of many prohormone molecules typically requires endoproteolytic cleavage C-terminal of dibasic residues, such as the K¹⁴¹R¹⁴²↓ site N-terminal of the α-melanocyte stimulating hormone (α-MSH) sequence in Silurana tropicalis proopiomelanocortin (POMC). In order to determine the absolute requirement of basic amino acid residues in the cleavage process, site-directed mutagenesis was employed to substitute alanine for the wild-type residues in the frog POMC open reading frame. Specifically, the following underlined residues were individually targeted for alanine substitution: R ¹³⁷ Q ¹³⁸ E ¹³⁹ N ¹⁴⁰ K ¹⁴¹ R ¹⁴²↓. The mouse pancreatic αTC1.9 cell …

Role Of Pro-Apoptotic Bcl2-Homology-3 Domain (Bh3)-Only Proteins In The Mutant Sod1 Mouse Model Of Als, Anna George Andrianakos

Electronic Theses and Dissertations

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease involving rapid degeneration of motor neurons in the spinal cord and retraction of their axonal projections to the neuromuscular junctions. Several known mutations linked to some familial cases of ALS have been linked to mutations in Cu/Zn superoxide dismutase (SOD1), resulting in mitochondrial oxidative stress and intrinsic apoptosis. Transgenic mice expressing a G93A mutant of SOD1 provide an in vivo model to investigate motor neuron death during disease progression. The principal regulators of intrinsic apoptosis are the Bcl-2 family proteins. While some members of this family are pro-survival, the Bcl-2 homology-3 …

Assessing Crosstalk Between Calcium And Camp In Pancreatic Alpha Cells, Julianne Michelle Imseis

Electronic Theses and Dissertations

Blood glucose levels are regulated by the interactions between insulin and glucagon. Pancreatic alpha cells release glucagon in response to a drop in blood sugar. Pancreatic alpha cells appear to be regulated through multiple signaling pathways. One potential method of regulation occurs from α-MSH or epinephrine stimulated production of the intracellular messenger cyclic AMP. Exocytosis of dense core vesicles containing glucagon ultimately requires an increase in cytosolic calcium. This study focused on the link between cyclic AMP and calcium in pancreatic alpha cells which allows alpha cell regulation. Interactions between calcium and cyclic AMP in pancreatic alpha cells consists of …

Minimotif Miner 2nd Release: A Database And Web System For Motif Search, Sanguthevar Rajasekaran, Sudha Balla, Patrick R. Gradie, Michael R. Gryk, Krishna Kadaveru, Vamsi Kundeti, Mark W. Maciejewski, Tian Mi, Nicholas Rubino, Jay Vyas, Martin R. Schiller

Life Sciences Faculty Research

Minimotif Miner (MnM) consists of a minimotif database and a web-based application that enables prediction of motif-based functions in user-supplied protein queries. We have revised MnM by expanding the database more than 10-fold to approximately 5000 motifs and standardized the motif function definitions. The web-application user interface has been redeveloped with new features including improved navigation, screencast-driven help, support for alias names and expanded SNP analysis. A sample analysis of prion shows how MnM 2 can be used.

Amino Acid Residues Implicated In The Interaction Of Melanocortin Ligands And Their Receptors: A Study Of Mc2r Selectivity, Kristopher D. Veo

Electronic Theses and Dissertations

Melanocortin receptor ligand selectivity has been a question not easily answered. The inability to functionally express melanocortin 2 receptor (MC2R) has inhibited the study of why MC2R is only stimulated by ACTH, a melanocortin hormone. With the recent discovery of the MC2R accessory protein (MRAP), creating a heterologous system is now feasible. Using a general cell line like CHO-K1 cells, which do not express endogenous MCRs, we were able to create a heterologous expression system and test the selectivity of MC2R using analog variants of ACTH(1-24). Our results indicate an amino acid requirement in the C-terminal portion of ACTH(1-24) for …

Erk Activation Requires Cam Kinases In Mcf-7 Breast Cancer Cells, John M. Schmitt, Ellen Abell

Faculty Publications - Department of Biological & Molecular Science

Abstract A key signaling pathway involved in regulating cell growth and proliferation throughout the body is the ERK signaling pathway. ERK is activated via numerous pathways including intracellular calcium release downstream of G-Protein Coupled Receptors (GPCRs). Carbachol, a GPCR-agonist, both increases intracellular calcium and ERK activation in MCF-7 cells. ERK activation and control of cell growth may act through the transcription factor Elk-1. Our goal was to elucidate the specific proteins and kinases upstream of ERK in MCF-7 cells treated with carbachol. Secondly, we wanted to investigate the potential involvement of Elk-1 downstream of ERK. Carbachol treatment of MCF-7 cells …

Cam Kinase Kinase Control Of Prostate Cancer Cell Survival, John M. Schmitt, Brendon Hart

Faculty Publications - Department of Biological & Molecular Science

Abstract AKT has been implicated in promoting cell survival within certain cells. Current research has shown that hormones that increase the concentration of intracellular calcium can activate AKT that in turn leads to cancer cell survival. Interestingly, LNCaP cells express the M3-subtype of GPCR's that may couple to increases in intracellular calcium and activation of the Ca2+/Calmodulin-dependent Protein Kinases (CaM Ks). Specifically CaM Kinase Kinase (CaM KK) phosphorylates its direct substrates CaM Kinase I, CaM Kinase IV, and AKT. AKT promotes cell survival through phosphorylation of its target BAD that prevents caspase activation. Our goals were to examine the mechanism …

Arbuscular Mycorrhizal Fungi: Factors Affecting Soil Organic Matter Storage, Carl Lee Rosier

Graduate Student Theses, Dissertations, & Professional Papers

Arbuscular mycorrhizal fungi (AMF) are soil fungi forming symbiotic associations with majority of land plants. AMF alter soil organic matter (SOM) directly through stabilization of soil aggregates and indirectly providing a path in which plant fixed C02 is transferred below-ground. Understanding contributions of AMF to SOM via protein production and stabilization of soil aggregates will greatly aid our understanding of soil carbon sequestration, nutrient cycling and mitigation of soil erosion. The work presented in chapter 2 challenges the glomalin extraction process and assesses the accuracy of the Bradford and monoclonal-antibody ELISA detection methods. My results clarify the contribution of glomalin …

Comparative Efficacy Of Bioud To Other Commercially Available Arthropod Repellants Against The Ticks Amblyomma Americanum And Dermacentor Variabilis On Cotton Cloth, Brooke W. Bissinger, Jiwei Zhu, Charles S. Apperson, Daniel E. Sonenshine, D. Wesley Watson, R. Michael Roe

Biological Sciences Faculty Publications

BioUD is an arthropod repellent that contains the active ingredient 2-undecanone originally derived from wild tomato plants. Repellency of BioUD was compared with five commercially available arthropod repellents against the ticks Amblyomma americanum (L.) and Dermacentor variabilis Say in two-choice bioassays on treated versus untreated cotton cheesecloth. Overall mean percentage repellency against both species was greatest for and did not differ significantly between BioUD (7.75% 2-undecanone) and products containing 98.1% DEET, 19.6% IR3535, and 30% oil of lemon eucalyptus. Products containing 5% and 15% Picaridin and 0.5% permethrin were also repellent compared with untreated controls but to a lesser degree …

Substrate And Regulation Of Mitochondrial Μ-Calpain, Aashish Joshi

University of Kentucky Doctoral Dissertations

μ -Calpain is localized to the mitochondrial intermembrane space. Apoptosisinducing factor (AIF), which executes caspase-independent cell death, is also localized to the mitochondrial intermembrane space. Following processing at the N-terminus, AIF becomes truncated (tAIF) and is released from mitochondria. The protease responsible for AIF processing has not been established. The same submitochondrial localization of mitochondrial μ-calpain and AIF gives support to the hypothesis that mitochondrial μ-calpain may be responsible for processing AIF. Atractyloside-induced tAIF release in rat liver mitochondria was inhibited by cysteine protease inhibitor MDL28170, but not by calpain inhibitors PD150606 or calpastatin. Moreover, μ-calpain immunoreactivity was difficult to …

Biochemical Characterization Of Two Yeast Paralogous Proteins Mth1 And Std1, Satish Pasula

Dissertations

Glucose is the most abundant monosaccharide and preferred carbon and energy source for most cells. Many organisms have evolved sophisticated means to sense glucose and respond to it appropriately. The budding yeast, Saccharomyces cerevisiae senses glucose through two transmembrane proteins, Snf3 and Rgt2. In the presence of extracellular glucose Snf3 and Rgt2 generate an intracellular signal that leads to the degradation of Mthl and Stdl, thereby inducing the expression of hexose transporter genes (EXT) by inhibiting the function of Rgtl, a transcriptional repressor of HXT genes. Mthl and Stdl are degraded via the Yckl/2 Kinase-SCFGrrl-26S proteasome pathway triggered by the …

Involvement Of Methyltransferases Enzymes During The Energy Metabolism Of The Anaerobic Archaeon Methanosarcina Semesiae Sp. Nov., Tj Lyimo

Tanzania Journal of Science

The methyl group transfer from dimethylsulfide (DMS), trimethylamine and methanol to 2-mercaptoethanesulfonic acid (coenzyme M) were investigated from cell extracts of Methanosarcina semesiae sp. nov. to evaluate whether the enzyme systems involved were constitutive or inductive. The extracts from cells grown on DMS exhibited methanogenic activity exclusively with DMS and methanethiol. Likewise when cells were pre-grown on trimethylamine or methanol the extracts only produced methane from the respective metaboilic substrate. Dimethylsulfide:methyl-coenzyme M transferase activity was dependent on ATP, but hydrogen did not stimulate activity. The fact that ATP could be replaced by the reductant Ti(III)-citrate indicates that reductive activation of …

Molecular Characterisation Of A Bovine-Like Rotavirus Detected From A Giraffe, Emily Mulherin, Jill Bryan, Marijke Beltman, Luke O'Grady, Eugene Pidgeon, Lucie Garon, Andrew Lloyd, John Bainbridge, Helen O'Shea, Paul Whyte, Séamus Fanning

Department of Biological Sciences Publications

Background

Rotavirus (RV), is a member of the Reoviridae family and an important etiological agent of acute viral gastroenteritis in the young. Rotaviruses have a wide host range infecting a broad range of animal species, however little is known about rotavirus infection in exotic animals. In this paper we report the first characterisation of a RV strain from a giraffe calf.

Results

This report describes the identification and detailed molecular characterisation of a rotavirus strain detected from a 14-day-old Giraffe (Giraffa camelopardalis), presenting with acute diarrhea. The RV strain detected from the giraffe was characterized molecularly as G10P[11]. …

Global Analysis Of Gene Expression Changes During Retinoic Acid-Induced Growth Arrest And Differentiation Of Melanoma: Comparison To Differentially Expressed Genes In Melanocytes Vs Melanoma, Mary H. Estler, Goran Boskovic, James Denvir, Sarah Miles, Donald A. Primerano, Richard M. Niles

Biochemistry and Microbiology

BACKGROUND: The incidence of malignant melanoma has significantly increased over the last decade. Some of these malignancies are susceptible to the growth inhibitory and pro-differentiating effects of all-trans-retinoic acid (RA). The molecular changes responsible for the biological activity of RA in melanoma are not well understood.

RESULTS: In an analysis of sequential global gene expression changes during a 4-48 h RA treatment of B16 mouse melanoma cells, we found that RA increased the expression of 757 genes and decreased the expression of 737 genes. We also compared the gene expression profile (no RA treatment) between non-malignant melan-a mouse melanocytes and …

Sex- And Gonad-Affecting Scent Compounds And 3 Male Pheromones In The Rat, Jian-Xu Zhang, Lixing Sun, Jin-Hua Zhang, Zhi-Yong Feng

All Faculty Scholarship for the College of the Sciences

This study was aimed at identifying sex pheromones of the rat (Rattus norvegicus). We characterized the volatiles and semivolatiles of rat preputial gland and voided urine by using gas chromatography–mass spectrometry (GC–MS) and quantified them by their GC areas (abundances) and percentage of GC areas (relative abundances). Although all the compounds other than 4-heptanone and phenol detected were shared by males and females, the quantities for some of these sex-common compounds exhibited sexual dimorphism and decreased with gonadectomy. Thus, these compounds might be sex pheromones. Among them, squalene from preputial glands and 2-heptanone and 4-ethyl phenol from urine …

Computational Design Of Orthogonal Ribosomes, Lon Chubiz, Christopher Rao

Biology Department Faculty Works

Orthogonal ribosomes (o-ribosomes), also known as specialized ribosomes, are able to selectively translate mRNA not recognized by host ribosomes. As a result, they are powerful tools for investigating translational regulation and probing ribosome structure. To date, efforts directed towards engineering o-ribosomes have involved random mutagenesisbased approaches. As an alternative, we present here a computational method for rationally designing o-ribosomes in bacteria. Working under the assumption that base-pair interactions between the 16S rRNA and mRNA serve as the primary mode for ribosome binding and translational initiation, the algorithm enumerates all possible extended recognition sequences for 16S rRNA and then chooses those …

Promoter Library Designed For Fine-Tuned Gene Expression In Pichia Pastoris, Franz S. Hartner, Claudia Ruth, David Langenegger, Sabrina N. Johnson, Petr Hyka, Geoff P. Lin-Cereghino, Joan Lin-Cereghino, Karin Kovar, James Cregg, Anton Glieder

College of the Pacific Faculty Articles

Although frequently used as protein production host, there is only a limited set of promoters available to drive the expression of recombinant proteins in Pichia pastoris. Fine-tuning of gene expression is often needed to maximize product yield and quality. However, for efficient knowledge-based engineering, a better understanding of promoter function is indispensable. Consequently, we created a promoter library by deletion and duplication of putative transcription factor-binding sites within the AOX1 promoter (P_AOX1) sequence. This first library initially spanned an activity range between ∼6% and >160% of the wild-type promoter activity. After characterization of the promoter library employing …

Detritus Type Alters The Outcome Of Interspecific Competition Between Aedes Aegypti And Aedes Albopictus (Diptera : Culicidae), Ebony G. Murrell, Steven A. Juliano

Faculty Publications – Biological Sciences

Many studies of interspecific competition between Aedes albopictus (Skuse) and Aedes aegypti (L.) (Diptera: Culicidae) larvae show that Ae. albopictus are superior resource competitors to Ae. aegypti. Single-species studies indicate that growth and survival of Ae. albopictus and Ae. aegypti larvae are affected by the type of detritus present in containers, which presumably affects the amount and quality of microorganisms that the mosquito larvae consume. We tested whether different detritus types alter the intensity of larval competition by raising 10 different density/species combinations of Ae. albopictus and Ae. aegypti larvae under standard laboratory conditions, with one of four detritus types …

Intracellular Levels Of Centrin2, Xpc, & Hhr23b Increase After Dna Damage, Nicholas A. Rowland

Honors Capstones

Capstone submitted as a graduate requirement for the BSU Honors Program.

A Watershed Analysis Of Pattee Pond: Implications For Water Quality And Land Use Management, Problems In Environmental Science Course (Biology 493), Colby College, Colby Environmental Assessment Team, Colby College

Colby College Watershed Study: Pattee Pond (2008, 1992)

The Colby Environmental Assessment Team (CEAT) investigated the impact of land use patterns on the water quality of Pattee Pond in Winslow, Maine, during the summer and fall of 2008. Physical, chemical, and biological characteristics of water quality were analyzed to evaluate the current health of the lake. Data collected were compared with previous studies conducted by the Maine Department of Environmental Protection (DEP) and CEAT to examine changes in water quality and land use over time. Trends in Pattee Pond water quality suggest an improvement since the 1970s. However, the lake is still experiencing algal blooms resulting from phosphorus …

Sustainability In A Time Of Climate Change: Developing An Intensive Research Framework For The Platte River Basin And The High Plains Proceedings From The 2008 Climate Change Workshop, Monica Norby, Ashley Washburn

Office of Research and Economic Development: Publications

Proceedings from the 2008 Climate change Workshop, Nebraska Sandhills, May 19-22, 2008. Hosted by the University of Nebraska–Lincoln and the U.S. Geological Survey

Carbachol Regulation Of Akt In Lncap Prostate Cancer Cells, John M. Schmitt, Luke Fletcher

Faculty Publications - Department of Biological & Molecular Science

Abstract Hormones and agonists that enhance cell survival through binding specific G Protein-Coupled Receptors (GPCRs) are of particular interest in cancer cell survival. LNCaP cells have been shown to express muscarinic cholinergic receptors that are responsive to the agonist, carbachol. In addition, LNCaP cells specifically express the M3-subtype of GPCR’s that may couple carbachol to Gq, increases in intracellular calcium, and activation of the intracelluar Ca2+/calmodulin-dependent protein kinases (CaM Ks). The CaM Kinase family of proteins includes CaM KII, CaM Kinase Kinase (CaM KK) and its substrates CaM KIV, CaM KI and the protein kinase AKT. AKT is an anti-apoptotic …

M3-Muscarinic Receptor Activation Of Erk And Cell Growth Requires Calcium/Calmodulin-Dependent Protein Kinases In Mcf-7 Cells, John M. Schmitt, Ellen Abell, Andrea Wagner

Faculty Publications - Department of Biological & Molecular Science

Abstract The extracellular signal-regulated protein kinase (ERK) signaling pathway is found in diverse cells throughout the human body. ERK activation has been implicated in breast cancer cell growth and proliferation. Studies have shown that ERK is activated by carbachol, a G Protein-Coupled Receptor (GPCR) agonist, which increases intracellular calcium in MCF-7 cells. The calcium/calmodulin-dependent protein kinase (CaM K) family of proteins including CaM KK, CaM KI, and CaM KII can be activated by increased intracellular calcium. Our goal was to determine whether CaM Ks may be responsible for ERK activation and cell proliferation in carbachol-treated MCF-7 cells and evaluate which …

Modulation Of Fast-Inactivating Potassium Channels By Accessory Subunits And Fatty Acids : A Role For Calcium?, Michelle M. Drzewiecki

Honors Theses

Potassium channel interacting proteins (KChIPs) co-assemble with Kv4 a subunits to form native complexes that encode considerable components of neuronal somatodendritic A- type K+current (Is,J in neurons, and transient outward current (ho) in cardiac myocytes. The binding of KChIPs to the cytoplasmic N-termini of Kv4 a subunits enhances surface expression by facilitating intracellular trafficking, stimulates subunit assembly, and regulates the functional gating properties of Kv4 channels. KChIPs, like many other neuronal calcium sensor proteins, contain four EF-hand calcium binding sites; two of which (EF-3 and 4) have particularly high affinity for calcium. Because Ca2+ is an important signaling molecule for …

Use Of Trees By The Texas Ratsnake (Elaphe Obsoleta) In Eastern Texas, Josh B. Pierce, Robert R. Fleet, Lance D. Mcbrayer, D. Craig Rudolph

Biology: Faculty Publications

We present information on the use of trees by Elaphe obsoleta (Texas Ratsnake) in a mesic pine-hardwood forest in eastern Texas. Using radiotelemetry, seven snakes (3 females, 4 males) were relocated a total of 363 times from April 2004 to May 2005, resulting in 201 unique locations. Snakes selected trees containing cavities and used hardwoods and snags for a combined 95% of arboreal locations. Texas Ratsnake arboreal activity peaked during July and August, well after the peak of avian breeding activity, suggesting arboreal activity involves factors other than avian predation.