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Biochemical Aspects Of The Thermal Sensitivity And Energy Balance Of Polar, Tropical And Subtropical Teleosts, Eloy Martinez Jan 2013

Biochemical Aspects Of The Thermal Sensitivity And Energy Balance Of Polar, Tropical And Subtropical Teleosts, Eloy Martinez

Graduate Theses and Dissertations

The maintenance of a functional energy balance in ectothermic fauna could be challenging in a thermally disparate environment. Biochemical adaptations at the enzyme and membrane levels allows for a set compensatory mechanism that allow the individual to maintain an energetic surplus, thus allocating energy for growth and reproduction. The present work describes how the energetic machinery in the cell, particularly the mitochondrion, could be affected by temperature changes. More specifically, this work aimed to determine how environmental temperature affects the mitochondria energetic performance of fishes from disparate thermal regimes.

Mitochondrial ATP production efficiency was evaluated in fishes from polar, tropical ...


Regulation Of The Tumor Suppresser P53 And Survivin By Ras And Ral Gtpases:Implications For Malignant Transformation, Awet G. Tecleab Jan 2013

Regulation Of The Tumor Suppresser P53 And Survivin By Ras And Ral Gtpases:Implications For Malignant Transformation, Awet G. Tecleab

Graduate Theses and Dissertations

Abstract

Although the critical role of the small GTPases Ras and Ral in oncogenesis has been well documented, much remains to be investigated about the molecular mechanism by which these GTPases regulate malignant transformation. The work under this thesis made two major contributions to this field. The first is the discovery that K-Ras, RalA and/or RalB are required for the maintenance of the high levels of the anti-apoptotic protein survivin in some human cancer cells, and the second is the demonstration that down regulation of K-Ras, RalA and/or RalB, but not Raf-1 or Akt1/2, stabilizes the tumor ...


Molecular Regulation Of Il-2 Inducible T-Cell Kinase (Itk) And The Tec Kinases: A Combined Experimental And Computational Study, With Emphasis On The N-Terminal Pleckstrin Homology Domain, Scott Edward Boyken Jan 2013

Molecular Regulation Of Il-2 Inducible T-Cell Kinase (Itk) And The Tec Kinases: A Combined Experimental And Computational Study, With Emphasis On The N-Terminal Pleckstrin Homology Domain, Scott Edward Boyken

Graduate Theses and Dissertations

This dissertation investigates the complex regulation of IL-2 inducible T-cell kinase (Itk) and its related Tec family kinase members, using a combination of experimental and computational approaches. Tec kinases are expressed primarily in hematopoietic cells and regulate key signaling events in lymphocyte activation, differentiation, and development. Itk functions in immune response, fine-tuning signaling downstream of the T-cell Receptor (TCR), and is a putative drug target for allergies and autoimmune disease. Btk, a Tec kinase found in B-cells, carries out a similar role to that of Itk, only downstream of B-cell Receptor (BCR) signaling. The chapters of this dissertation focus on ...


Design Of Novel Nano-Carriers For Multi-Enzyme Co-Localization, Feng Jia Jan 2013

Design Of Novel Nano-Carriers For Multi-Enzyme Co-Localization, Feng Jia

Graduate Theses and Dissertations

The widely existing MECs in Nature have inspired researchers to design synthetic analogs to promote the overall catalytic efficiency in vitro by co-localizing multiple enzymes to mimic the MECs' unique functionalities. A number of efforts have been devoted to designing synthetic MECs in the past couples of decades. This thesis work has focused on developing novel strategies based on enzyme immobilization to design nano-carriers for multi-enzyme co-localization to realize kinetics enhancement and strong control of spatial arrangement of the enzymes. Three distinct approaches have been designed using different attachment methods and platforms.

First, the multifunctional polystyrene nanoparticles were designed for ...


Development Of Strategies For Synthesis Of Branched Caenorhabditis Elegans-Associated Oligosaccharides And Of Photo-Activated Lectin Capture Reagents, Randy Alan Benedict Jan 2013

Development Of Strategies For Synthesis Of Branched Caenorhabditis Elegans-Associated Oligosaccharides And Of Photo-Activated Lectin Capture Reagents, Randy Alan Benedict

Graduate Theses and Dissertations

The development and investigation of a new affinity proteomics probe and methodology are presented in this dissertation. A trifunctional affinity proteomics probe (TAPP) was envisioned and prepared to serve as a lectin capture reagent. The TAPP consists of a solid support to serve as a handle for captured protein as well as scaffold for multivalency, a saccharide of interest to fish for an unknown or elicit a suspected specific interaction, and a photoactivated cross-linking moiety to covalently capture binding proteins. The perfluorinated phenylazide cross-linker was shown to exhibit better performance relative to the parent phenylazide with respect to sensitivity and ...


Investigation Of The Process Of Snare Mediated Synaptic Vesicle Exocytosis, Ying Lai Jan 2013

Investigation Of The Process Of Snare Mediated Synaptic Vesicle Exocytosis, Ying Lai

Graduate Theses and Dissertations

Neurotransmitter release in synapses underlies major brain functions such as cognition, emotion, and memory. The precise temporal control of the release is essential for healthy brain activities. SNAREs are known to be the core fusion machinery in neuro-exocytotic pathways. SNARE complex formation in the chasm of two membranes is mediated by cognate coiled-coil motifs on v- and t-SNAREs: One such motif from t-SNARE syntaxin 1A, two from t-SNARE SNAP-25, and another from v-SNARE VAMP2 all form a parallel four-stranded coiled coil that brings about the apposition of two membranes. However, The SNARE proteins do not have the required regulatory function ...


Characterization Of The Putative Catalytic Domain Of Two Xyloglucan Xylosyltransferases (Xxts) Involved In The Biosynthesis Of Xyloglucan In Arabidopsis Thaliana, Rachel Ann Morris Jan 2013

Characterization Of The Putative Catalytic Domain Of Two Xyloglucan Xylosyltransferases (Xxts) Involved In The Biosynthesis Of Xyloglucan In Arabidopsis Thaliana, Rachel Ann Morris

Graduate Theses and Dissertations

Xyloglucan is the primary hemicellulosic component of the primary cell wall in dicotyledonous plants. It is characterized by a beta-(1-4)-D-glucan backbone that is substituted in a regular pattern with alpha-(1-6)-D-xylose, and can be substituted further with beta-(1-2)-D-galactose or beta-(1-2)-D-galactose-alpha(1-2)-L-fucose disaccharide. Three xyloglucan xylosyltransferases (XXTs), XXT1, XXT2, and XXT5, are responsible for the D-xylosylation of the beta-(1-4)-D-glucan backbone in Arabidopsis thaliana. They are Golgi-localized type II transmembrane proteins that have putative DXD catalytic centers. All have been shown to interact in a protein complex, XXT2 and XXT5 showing the ...


Allosteric Regulation Of Mammalian Fructose-1,6-Bisphosphatase, Yang Gao Jan 2013

Allosteric Regulation Of Mammalian Fructose-1,6-Bisphosphatase, Yang Gao

Graduate Theses and Dissertations

Fructose-1, 6-bisphosphate (D-fructose-1, 6-bisphosphate 1-phosphohydrolase; EC 3. 1. 3; FBPase) is an essential regulatory enzyme in gluconeogenesis and has long been considered as a drug target towards type II diabete. In mammalian, the activity of FBPase is regulated by AMP and fructose 2,6-bisphosphate (Fru-2,6-P2). AMP is an allosteric inhibitor that binds to FBPase with positive coopertivity, and Fru-2,6-P2 is an active site inhibitor which is up-regulated by hormone. Despite the 30 Å distance between their binding sites, both of AMP and Fru-2,6-P2 transform FBPase from active R-state to inactive T-state. Large conformational rearrangements are coupled to ...


Investigation Of The Arabidopsis Nonhost Resistance Mechanism Against The Soybean Pathogen, Phytophthora Sojae, Rishi Sumit Jan 2013

Investigation Of The Arabidopsis Nonhost Resistance Mechanism Against The Soybean Pathogen, Phytophthora Sojae, Rishi Sumit

Graduate Theses and Dissertations

Nonhost resistance (NHR) provides immunity to all members of a plant species against all isolates of a microorganism that is pathogenic to other plant species. Three Arabidopsis thaliana PEN (penetration deficient) genes, PEN1, 2 and 3 have been shown to provide prehaustorial NHR against the barley pathogen Blumeria graminis f. sp. hordei. Arabidopsis pen1-1 mutant is penetrated by the hemibiotrophic oomycete pathogen, Phytophthora sojae that causes root and stem rot disease in soybean. The P. sojae susceptible (pss) 1 mutant is infected by both P. sojae and the hemibiotrophic fungal pathogen, Fusarium virguliforme that causes sudden death syndrome in soybean ...


Sustainable Chemistry, Synthesis And Structure-Activity Relationship Studies Of Biologically Important Small Molecules, Gerald R. Pollock, Iii Jan 2013

Sustainable Chemistry, Synthesis And Structure-Activity Relationship Studies Of Biologically Important Small Molecules, Gerald R. Pollock, Iii

Graduate Theses and Dissertations

With growing concerns for the environment, dwindling amounts of fossil fuels available for worldwide consumption and more resistant strains of diseases and infections becoming a reality, organic chemistry plays a vital role in the improvement and sustainability of modern society. Industrial chemicals from renewable sources has become an important area of research in recent years, while the synthesis and study of biologically active small molecules and their analogues leads to advancements which can help improve overall human health.

In this dissertation, we explore methods to create important industrial molecules from biorenewable sources, extend a novel cyclization to make heterocyclic compounds ...


Exploring And Understanding The Substrate Diversity Of B-Ketoacyl-Acp Synthase Iii Enzymes, Shivani Garg Jan 2013

Exploring And Understanding The Substrate Diversity Of B-Ketoacyl-Acp Synthase Iii Enzymes, Shivani Garg

Graduate Theses and Dissertations

Fatty acids are an integral part of the lipid membrane of bacteria and plants, and the enzyme that initiates fatty acid synthesis, β-ketoacyl-ACP Synthase III (KASIII), is one of the most diverse enzymes found in nature. KASIII exhibits vast diversity in its substrate specificity. By virtue of its substrate specificity, KASIII determines the fatty acid profile of the organism by dictating the nature of the ω-end of the fatty acids. For example, KASIIIs from many bacteria (e.g. Escherichia coli) accept straight chain acyl-CoAs resulting in production of straight chain fatty acids and are, therefore, narrow in substrate specificity. In ...


Structure And Function Of Class One Non-Symbiotic Plant Hemoglobins, Ryan Thomas Sturms Jan 2013

Structure And Function Of Class One Non-Symbiotic Plant Hemoglobins, Ryan Thomas Sturms

Graduate Theses and Dissertations

Plants contain at least three kinds of hemoglobins. Those plants that carry out symbiotic nitrogen fixation use oxygen transport hemoglobins to deliver oxygen to the aerobic nitrogen fixing bacteria in their roots. The functions of other plant hemoglobins are not yet known with confidence, but are thought to also have roles in nitrogen metabolism. This dissertation examines plant hemoglobin structure and function in two distinct classes: oxygen transport hemoglobins and what we believe to be hemoglobins that function as dissimilatory nitrite reductases.

The capacity for oxygen transport arose twice independently in two distinct phylogenetic classes of plant hemoglobins. From "Class ...


Structural And Functional Study Of Snare Machinery In Neurotransmitter Release, Shuang Song Jan 2013

Structural And Functional Study Of Snare Machinery In Neurotransmitter Release, Shuang Song

Graduate Theses and Dissertations

Synaptic neurotransmitter release is the most critical communication process for the connections between neurons and between neurons and target cells. SNAREs (soluble N-ethylmaleimide sensitive factor attachment protein receptors) are believed to be highly involved in docking and fusion of synaptic vesicles to the pre-synaptic plasma membrane. In vivo, synaptic vesicle exocytosis is a regulated and extremely rapid process. Numerous regulatory proteins are also required to achieve the fast speed and Ca2+ dependency of synaptic neurotransmitter release.

Our research mainly focuses on investigating the mechanism and protein structural basis of SNARE mediated membrane fusion. Site-directed spin labeling (SDSL) and electron paramagnetic ...


Structural Basis For Ternary Complex Formation Between Tau, Hsp90, And Fkbp51, Alexander Steven Barrett Jan 2013

Structural Basis For Ternary Complex Formation Between Tau, Hsp90, And Fkbp51, Alexander Steven Barrett

Graduate Theses and Dissertations

The accumulation of the microtubule associated protein tau has been implicated in several neurological disorders; however, its interaction with chaperones along its normal degradation pathway remains largely uncharacterized at single residue resolution. In this study, nuclear magnetic resonance (NMR) spectroscopy was used to probe the interaction between tau, the molecular chaperone Hsp90, and the immunophilin FKBP51. Resonance intensity changes were observed for specific residues in the heteronuclear single quantum coherence (HSQC) spectra of 15N-labeled tau in the presence of Hsp90 and/or FKBP51. Analysis of the HSQC spectra identified the two hydrophobic hexapeptide motifs located at residues V275 - K280 and ...


Structure, Dynamics, And Evolution Of The Intrinsically Disordered P53 Transactivation Domain, Wade Michael Borcherds Jan 2013

Structure, Dynamics, And Evolution Of The Intrinsically Disordered P53 Transactivation Domain, Wade Michael Borcherds

Graduate Theses and Dissertations

in numerous disease states, including cancers and neurodegenerative diseases. All proteins are dynamic in nature, occupying a range of conformational flexibilities. This inherent flexibility is required for their function, with ordered proteins and IDPs representing the least flexible, and most flexible, respectively. As such IDPs possess little to no stable tertiary or secondary structure, they instead form broad ensembles of heterogeneous structures, which fluctuate over multiple time scales. Although IDPs often lack stable secondary structure they can assume a more stable structure in the presence of their binding partners in a coupled folding binding reaction.

The phenomenon of the dynamic ...


Immobilization And Characterization Of Physisorbed Antibody Films Using Pneumatic Spray As Deposition Technique, Jhon J. Figueroa Jan 2013

Immobilization And Characterization Of Physisorbed Antibody Films Using Pneumatic Spray As Deposition Technique, Jhon J. Figueroa

Graduate Theses and Dissertations

The immobilization of antibodies on silica surfaces has been a wide and common practice via cross-linking with the formation of covalent bonds between surface and antibody. The formation of antibody thin films on solid surfaces using pneumatic spray (PS) as the deposition technique and the analysis of the surface morphology of these films were investigated during this study. The pneumatic spray method was compared with the covalent bonding method Avidin-Biotin Bridge (ABB). The intensities and capture efficiency tests showed similar results for both techniques with a lower signal-to-noise ratio (SNR) for the PS deposited films. Specificity tests suggested that the ...


Optical Investigations Of Neurohypophysial Excitability And Amyloid Fibril Formation, Joseph Leo Foley Jan 2013

Optical Investigations Of Neurohypophysial Excitability And Amyloid Fibril Formation, Joseph Leo Foley

Graduate Theses and Dissertations

This dissertation describes the work done on two distinct projects. In the first part I sought to unravel the mechanisms that underlie the activity-dependent modulation of response in the excitation-secretion coupling of the neurohypophysis. In the second part, I optically monitored and analyzed the secondary structure changes accompanying amyloid fibril formation along multiple pathways, under both denaturing and near-physiological conditions.

Neuronal plasticity plays an important role in regulating various biological systems by modulating release of hormones or neurotransmitters. The changing response to the same stimulus, depending on the context and previous stimulation events, is also the basis of learning and ...


Reaction Enthalpy And Volume Profiles For Excited State Reactions Involving Electron Transfer And Proton-Coupled Electron Transfer, William Antonio Maza Jan 2013

Reaction Enthalpy And Volume Profiles For Excited State Reactions Involving Electron Transfer And Proton-Coupled Electron Transfer, William Antonio Maza

Graduate Theses and Dissertations

Electron transfer, ET, and proton-coupled electron transfer, PCET, reactions are central to biological reactions involving catalysis, energy conversion and energy storage. The movement of electrons and protons in either a sequential or concerted manner are coupled in a series of elementary reaction steps in respiration and photosynthesis to harvest and convert energy consumed in foodstuffs or by absorption of light into high energy chemi-cal bonds in the form of ATP. These electron transfer processes may be modulated by conformational dynamics within the protein matrix or at the protein-protein interface, the energetics of which are still not well understood. Photoacoustic calorimetry ...


Fabricating And Characterizing Physical Properties Of Electrospun Polypeptide-Based Nanofibers, Dhan Bahadur Khadka Jan 2013

Fabricating And Characterizing Physical Properties Of Electrospun Polypeptide-Based Nanofibers, Dhan Bahadur Khadka

Graduate Theses and Dissertations

This dissertation has aimed to fabricate polypeptide based biomaterial and characterize physical properties. Electrospinning is used as a tool for the sample fabrication. Project focused on determining the feasibility of electrospinning of certain synthetic polypeptides and certain elastin-like peptides from aqueous feedstocks and to characterize physical properties of polymer aqueous solution, cast film and spun fibers and fiber mats. The research involves peptide design, polymer electrospinning, fibers crosslinking, determining the extent of crosslinking, fibers protease degradation study, fibers stability and self-organization analysis, structure and composition determination by various spectroscopy and microscopy techniques and characterization of mechanical properties of individual suspended ...


Sirt1 Regulation Of The Heat Shock Response In An Hsf1-Dependent Manner And The Impact Of Caloric Restriction, Rachel Rene Raynes Jan 2013

Sirt1 Regulation Of The Heat Shock Response In An Hsf1-Dependent Manner And The Impact Of Caloric Restriction, Rachel Rene Raynes

Graduate Theses and Dissertations

The heat shock response (HSR) is the cell's molecular reaction to protein damaging stress and is critical in the management of denatured proteins. Activation of HSF1, the master transcriptional regulator of the HSR, results in the induction of molecular chaperones called heat shock proteins (HSPs). Transcription of hsp genes is promoted by the hyperphosphorylation of HSF1, while the attenuation of the HSR is regulated by a dual mechanism involving negative feedback inhibition from HSPs and acetylation at a critical lysine residue within the DNA binding domain of HSF1, which results in a loss of affinity for DNA. SIRT1 is ...


The Effects Of Heat Stress In Redox Balance And Inflammatory Signaling In Porcine Skeletal Muscle, Sandra Isabel Rosado Montilla Jan 2013

The Effects Of Heat Stress In Redox Balance And Inflammatory Signaling In Porcine Skeletal Muscle, Sandra Isabel Rosado Montilla

Graduate Theses and Dissertations

Heat stress (HS) is a major concern for human health as well as for the swine industry. In 2012, HS resulted in the largest number of weather related human fatalities in the US. Animal production is also compromised as HS leads to poor sow performance, decreased carcass value and increased veterinary costs. However, the molecular effects of HS on skeletal muscle are still unclear. This study aimed to determine the extent to which HS disrupted redox balance and initiated an inflammatory response in oxidative and glycolytic porcine skeletal muscle. Moreover, we quantified the contribution of reduced feed intake to the ...


Characterization Of The Arabidopsis Thaliana Acetyl-Coa Synthetase Putative Carboxylate Binding Pocket, Jason H. Hart Jan 2013

Characterization Of The Arabidopsis Thaliana Acetyl-Coa Synthetase Putative Carboxylate Binding Pocket, Jason H. Hart

Graduate Theses and Dissertations

Acetyl-CoA synthetase catalyzes the activation of acetate by the acetylation of the thiol group of Coenzyme A, while hydrolyzing ATP to AMP and pyrophosphate. The Arabidopsis thaliana acetyl-CoA synthetase (atACS) was compared to other acyl-CoA synthetases, and was computationally modeled on the available crystal structures of the Saccharomyces cerevisiae ACS1 and Salmonella enterica ACS. This allowed the identification of the residues that make up the putative carboxylate binding pocket residues. To further understand substrate selectivity and binding within the putative carboxylate binding pocket, selected residues were mutated to resemble the homologous residues in the Pseudomonas chlororaphis isobutyryl-CoA synthetase. Four residues ...


Determination And Quantification Of Surface Lipid Metabolites In Maize Silks: A Pathway Model For Surface Lipid Biosynthesis Based On Simultaneous Profiling Of Polar And Non-Polar Metabolites, Layton Andrew Peddicord Jan 2013

Determination And Quantification Of Surface Lipid Metabolites In Maize Silks: A Pathway Model For Surface Lipid Biosynthesis Based On Simultaneous Profiling Of Polar And Non-Polar Metabolites, Layton Andrew Peddicord

Graduate Theses and Dissertations

The elongated stigmas in maize (i.e. silks) are a vital organ for the reproductive success of a major U.S. agricultural crop. These silks possess an extractible coat of surface lipids that is predominantly composed of a complex array of hydrocarbons that function to protect the silks against abiotic and biotic stresses. While hydrocarbons are most likely produced from the conversion of fatty acids, the exact biochemical mechanisms for their synthesis remain unknown. Our team is leveraging genetic variation and morphological gradients during silk development to dissect the metabolic network responsible for surface lipid accumulation.

These metabolomic efforts require ...


Mass Spectrometry-Based Methods For The Detection And Characterization Of Protein-Tyrosine Nitration, Kent W. Seeley Jan 2013

Mass Spectrometry-Based Methods For The Detection And Characterization Of Protein-Tyrosine Nitration, Kent W. Seeley

Graduate Theses and Dissertations

Protein tyrosine nitration (PTN) is a posttranslational modification resulting from oxidative/nitrosative stress that has been implicated in a wide variety of disease states. Characterization of PTN is challenging due to several factors including its low abundance in a given proteome, preferential site modification, multiple target site proximity within unique peptide sequences, and analytical method and instrument limitations. Current analytical techniques are insufficiently sensitive to identify endogenous nitration sites without incorporation of either nitrotyrosine or target protein enrichment. However, enrichment proficiency can also be inadequate. Chemical derivatization of the nitro- moiety can be incomplete or result in undesirable byproduct formation ...


Phosphorylation Of Histone Deacetylase 6 Within Its C-Terminal Region By Extracellular Signal Regulated Kinase 1, Kendra Allana Williams Jan 2013

Phosphorylation Of Histone Deacetylase 6 Within Its C-Terminal Region By Extracellular Signal Regulated Kinase 1, Kendra Allana Williams

Graduate Theses and Dissertations

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Epigenetic Modifiers To Augment The Immunogenicity Of Chronic Lymphocytic Leukemia, Jason A. Dubovsky Jan 2013

Epigenetic Modifiers To Augment The Immunogenicity Of Chronic Lymphocytic Leukemia, Jason A. Dubovsky

Graduate Theses and Dissertations

Cancer cells employ a litany of immunosuppressive and immunevasive strategies to avoid detection and elimination by the various arms of the innate and adaptive immune system. Many hematologic and solid tumors progressively develop a specialized microenvironment which promotes tissue restructuring inflammation while masking the immune signature of the tumor cells themselves. Chronic lymphocytic leukemia, a malignancy of mature B lymphocytes must constantly balance on the precipice of immune recognition. A mature antigen presenting cell themselves, CLL clonal growth is dependent on the very interactions which, if effective, could potentially lead to their demise. To circumvent this, CLL clones set up ...


The Panicum Mosaic Virus-Like 3' Cap-Independent Translation Element: A Translation Enhancer That Functions In Mammalian Systems, Mariko Sada Peterson Jan 2013

The Panicum Mosaic Virus-Like 3' Cap-Independent Translation Element: A Translation Enhancer That Functions In Mammalian Systems, Mariko Sada Peterson

Graduate Theses and Dissertations

In canonical eukaryotic translation initiation, the recognition of mRNA by the translation machinery is facilitated through the binding of the 5' m7G cap structure to eukaryotic initiation factor 4E (eIF4E). This recognition is necessary for efficient translation of the message to occur. In contrast, some viruses lack the 5' cap structure but have alternative mechanisms to initiate translation and express the viral proteins necessary for replication. The positive-strand RNA plant viruses of the panico-, carmo-, umbra-, (1-3) and aureusviruses (4) contain an RNA structure in their 3' UTRs that binds eIF4E (2) and through communication with the 5' UTR, are ...


Building And Simulating Protein Machines, Ataur Rahim Katebi Jan 2013

Building And Simulating Protein Machines, Ataur Rahim Katebi

Graduate Theses and Dissertations

Glycolysis is a central metabolic pathway, present in almost all organisms, that produces energy. The pathway has been extensively investigated by biochemists. There is a significant body of structural and biochemical information about this pathway. The complete pathway is a ten step process. At each step, a specific chemical reaction is catalyzed by a specific enzyme. Fructose bisphosphate aldolase (FBA) and triosephosphate isomerase (TIM) catalyze the fourth and the fifth steps on the pathway.

This thesis investigates the possible substrate transfer mechanism between FBA and TIM. FBA cleaves its substrate, the six-carbon fructose-1,6-bisphosphate (FBP), into two three-carbon products - glyceraldehydes ...


Step-Specific Investigation Of Snare-Mediated Membrane Fusion, Sunae Kim Jan 2013

Step-Specific Investigation Of Snare-Mediated Membrane Fusion, Sunae Kim

Graduate Theses and Dissertations

Cholesterol is a major component of biological membranes and is known to affect vesicle fusion. However, the mechanism by which cholesterol modulates SNARE-dependent intracellular fusion is not well understood. First, using the fluorescence assay and employing dye-labeled SNAREs and the fluorescent lipids on yeast post golgi trafficking SNARE-mediated model membrane fusion, we dissected cholesterol effects on individual fusion steps including SNARE complex formation, hemifusion, pore formation, and pore dilation. At physiological high concentrations, cholesterol stimulated hemifusion as much as 30-fold, but its stimulatory effect diminished to 10- and 3-folds for subsequent pore formation and pore expansion at 40 mole%, respectively ...


Specificity Of The Cocaine Aptamer And Its Application In Measurement Of Cocaine And Metabolites In Homogeneous Samples For Forensic Analysis, Ashish Sachan Jan 2013

Specificity Of The Cocaine Aptamer And Its Application In Measurement Of Cocaine And Metabolites In Homogeneous Samples For Forensic Analysis, Ashish Sachan

Graduate Theses and Dissertations

Detection and quantification of molecules has an enormous importance and potential in the field of scientific development and basic discovery research. Advances in colorimetric techniques, fluorescent labeled probes and biosensor based technologies have resulted in the specific detection and precise quantification of analytes. The effective development of a particular detection method needs optimization in terms of overall speed, efficiency, and accuracy of the sensing system. Current biosensor approaches often rely on cumbersome procedural steps and reagent requirements. Along with the complexity of the assay, the size of the analytical instrument is a deterrent factor for on-site applications. To overcome such ...