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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Understanding Protein Motions By Computational Modeling And Statistical Approaches, Lei Yang Jan 2008

Understanding Protein Motions By Computational Modeling And Statistical Approaches, Lei Yang

Retrospective Theses and Dissertations

Because of its appealing simplicity, the elastic network model (ENM) has been widely accepted and applied to study many molecular motion problems, such as the molecular mechanisms of chaperonin GroEL-GroES function, allosteric changes in hemoglobin, ribosome motions, motor-protein motions, and conformational changes in general. In this dissertation, the ENM is employed to study various protein dynamics problems, and its validity is also examined by comparing with experimental data. First, we apply principal component analysis (PCA) to identify the essential protein motions from multiple structures (X-ray, NMR and MD) of the HIV-1 protease. We find significant similarities between the first few ...


Relatedness Of Haemophilus Parasuis Strains And Their Proteins' Possible Roles As Virulence Factors, Emilie Susanna Noel Zehr Jan 2008

Relatedness Of Haemophilus Parasuis Strains And Their Proteins' Possible Roles As Virulence Factors, Emilie Susanna Noel Zehr

Retrospective Theses and Dissertations

Haemophilus parasuis, the causative agent of Glasser's disease, is considered to be a remerging pathogen of swine in high-health status herds. Several molecular typing methods have been suggested as alternatives to serotyping, which failed to identify up to 30% of field isolates. This study compared random amplification of polymorphic DNA (RAPD) profiles and protein profiles of H. parasuis reference strains and field isolates as methods for identifying and grouping virulent and avirulent isolates. DNA and protein profiles of 15 reference strains and 31 field isolates of H. parasuis, analyzed with two computer-based similarity analyses, revealed unique and reproducible DNA ...


Molecular Interactions Of The Mammalian Intermediate Filament Protein Synemin With Cytoskeletal Proteins Present In Adhesion Sites, Ning Sun Jan 2008

Molecular Interactions Of The Mammalian Intermediate Filament Protein Synemin With Cytoskeletal Proteins Present In Adhesion Sites, Ning Sun

Retrospective Theses and Dissertations

The type VI intermediate filament (IF) protein synemin is a very large, unique member of the IF protein superfamily. Synemin associates with the major type III IF protein desmin and/or vimentin forming heteropolymeric intermediate filaments (IFs) within mammalian muscle cells. Previous studies in our lab showed that avian synemin interacts with vinculin and alpha-actinin, both of which are cytoskeletal proteins present in adhesion sites such as the focal adhesions within cells. Thus, synemin may link the heteropolymeric IFs to adhesion sites within mammalian muscle cells or within some non-muscle cells that express synemin. At least two isoforms of synemin ...


Functional Analysis Of The C-Terminal Domain In The Drosophila Jil-1 Histone H3 Kinase, Xiaomin Bao Jan 2008

Functional Analysis Of The C-Terminal Domain In The Drosophila Jil-1 Histone H3 Kinase, Xiaomin Bao

Retrospective Theses and Dissertations

Epigenetic regulation is an important process utilized by biological systems to control gene expression and organize development, with histone modification enzymes among the most important components in this regulatory network. The predominant interphase H3 Serine10 kinase in Drosophila is JIL-1, which plays an important role in regulating chromosome structure and modulating gene expression. The full-length JIL-1 protein is composed of four domains including a N-terminal domain (NTD), two kinase domains (KDI and KDII), and a unique C-terminal domain (CTD). In order to study the biological importance of these individual domains, a domain analysis research approach was taken and different functions ...


Dissection Of The Mitotic And Nuclear Functions Of Chromator, A Nuclear-Derived Spindle Matrix Component In Drosophila, Yun Ding Jan 2008

Dissection Of The Mitotic And Nuclear Functions Of Chromator, A Nuclear-Derived Spindle Matrix Component In Drosophila, Yun Ding

Retrospective Theses and Dissertations

A spindle matrix has long been proposed to serve as a stationary or elastic molecular matrix substrate for the organization and activities of microtubules and motors, based on the consideration of a mechanical and functional support for the stabilization of microtubule spindle during force generation from mitotic motors. Recently, the identification of four Drosophila proteins, Skeletor, Megator, EAST and Chromator has provided molecular evidence for the existence of this macromolecular matrix structure during mitosis. All of these four proteins have been shown to interact with each other within a protein complex and redistribute from the nucleus at interphase to form ...


Biochemical And Genetic Analyses Of Physical Associations Among Zea Mays Starch Biosynthetic Enzymes, Tracie Ann Hennen-Bierwagen Jan 2008

Biochemical And Genetic Analyses Of Physical Associations Among Zea Mays Starch Biosynthetic Enzymes, Tracie Ann Hennen-Bierwagen

Retrospective Theses and Dissertations

Mutations affecting specific starch biosynthetic enzymes commonly have pleiotropic effects on other enzymes in the same metabolic pathway. Such genetic evidence indicates functional relationships between components of the starch biosynthetic system including starch synthases (SS), starch branching enzymes (BE), and starch debranching enzymes (DBE), however, the molecular explanation for these functional interactions is not known. One possibility is that specific SSs, BEs, and/or DBEs associate physically with each other in multisubunit complexes. To test this hypothesis, this study sought to identify stable associations between SSI, SSIIa, SSIII, SBEI, SBEIIa, and SBEIIb from maize amyloplasts. Three separate detection methods, yeast ...


Development Of An Rna Aptamer To Pd173955n Through Selex, Samir Mehanovic Jan 2008

Development Of An Rna Aptamer To Pd173955n Through Selex, Samir Mehanovic

Retrospective Theses and Dissertations

With all the technology available to us, the causes of cancers are still puzzling researchers all over the world. Everyday different approaches are being tested to help cure various cancers. With the development of molecular biology, we are able to understand cancer more at molecular level, which helps us look at different methods to treat cancer patients.;The molecular basis for leukemia cancers are understood to a level where we are able to design treatments that specifically target proteins responsible for the disease. One of those proteins is the Bcr-Abl tyrosine kinase which is mostly responsible for chronic myelogenous leukemia ...


The Structure And Mechanism Of Saicar Synthetase, Nathaniel Daum Ginder Jan 2008

The Structure And Mechanism Of Saicar Synthetase, Nathaniel Daum Ginder

Retrospective Theses and Dissertations

SAICAR synthetase catalyzes the eighth step in bacterial de novo purine nucleotide biosynthesis: ATP + L-aspartate + CAIR → ADP + Pi + SAICAR Enzymes involved in nucleotide synthesis are common targets for chemotherapeutic and antimicrobial drugs. They work by reducing the rate of growth of rapidly proliferating cells, thus preventing tumor growth and microbial fecundity. L-Alanosine, an L-aspartate analogue that acts as a substrate for SAICAR synthetase, is a natural product with antiviral and antitumor activities. The product of the reaction is a potent inhibitor of adenylosuccinate synthetase and adenylosuccinate lyase and is responsible for L-alanosine toxicity. L-Alanosine may be effective as a chemotherapeutic ...


Studies Of Intermediates And Regulation In Snare-Mediated Membrane Fusion, Xiaobing Lu Jan 2008

Studies Of Intermediates And Regulation In Snare-Mediated Membrane Fusion, Xiaobing Lu

Retrospective Theses and Dissertations

At the synapse, neurotransmitters are released via Ca2+-triggered exocytotic fusion of synaptic vesicles with the presynaptic plasma membrane. The whole process is controlled by various proteins. SNAREs have been recognized as the key components that drive membrane fusion. In addition, many other proteins, such as Munc18/nSec1, Mun13, synaptotagmin, complexin, etc. are characterized to regulate synaptic transmission temporally and spatially.;The in vitro bulk fluorescence assay was applied to examine the kinetics of membrane fusion of liposomes mediated by recombinant neuronal SNAREs and led to the demonstration of hemifusion as an intermediate in the pathway. In order to monitor ...


Use Of Fourier Transform Infrared (Ftir) Spectroscopy To Determine The Health-Promoting Index (Hpi) Of Cow's Milk, Wendy Leanne Thomas Jan 2008

Use Of Fourier Transform Infrared (Ftir) Spectroscopy To Determine The Health-Promoting Index (Hpi) Of Cow's Milk, Wendy Leanne Thomas

Retrospective Theses and Dissertations

Fourier transform infrared (FTIR) spectroscopy steadily has been gaining popularity for the analysis of milk components including total fat, protein, lactose, and ketones. To date, there have been few descriptions of the use of FTIR spectroscopy for the determination of milk fatty acid composition.;The first objective of this thesis was to develop a method utilizing FTIR spectroscopy for prediction of the health-promoting index (HPI) of milk. The HPI is known to affect the relative softness of butter (Bobe et al., 2003); so, this model could be used to predict which cows' milk could be designated for generation of niche ...


Functional Analysis Of Paralogous Genes In Acetyl Coenzyme A Metabolism, Joel D. Schmidt Jan 2008

Functional Analysis Of Paralogous Genes In Acetyl Coenzyme A Metabolism, Joel D. Schmidt

Retrospective Theses and Dissertations

No abstract provided.


The Locations And Signaling Of H-Ras On Endosomes And Plasma Membrane, Xing Wang Jan 2008

The Locations And Signaling Of H-Ras On Endosomes And Plasma Membrane, Xing Wang

Retrospective Theses and Dissertations

No abstract provided.


The Roles Of Lcn2 In The Inflammatory Lung And Liver, Yinghua Liu Jan 2007

The Roles Of Lcn2 In The Inflammatory Lung And Liver, Yinghua Liu

Retrospective Theses and Dissertations

Uterocalin (Lcn2; also called 24p3, SIP24, or siderocalin and abbreviated as Utc) is an acute phase protein, for which the physiological function remains to be determined. Basal Utc expression is highest in the uterus, mammary gland and lung, which are three tissues with direct exposure to the diverse bacterial flora of the external environment. The demonstrated ability to bind siderophores is one means by which Utc can provide protection against pathogenic bacteria. However, its specificity for siderophores limits the ability of Utc to ward off pathogens. Epithelial tissues that interact with the external environment are exposed to many different nonpathogenic ...


Characterization Of Rns2, An S-Like Rnase In Arabidopsis Thaliana Suggests Role In Phosphate Recycling, Saralyn Ohanian Jan 2007

Characterization Of Rns2, An S-Like Rnase In Arabidopsis Thaliana Suggests Role In Phosphate Recycling, Saralyn Ohanian

Retrospective Theses and Dissertations

Ribonucleases play important biological roles within all organisms but many have not been well characterized. The family of T2 RNases is well-conserved throughout the kingdoms of life and yet their properties and functions are not well understood. RNS2 is one of five such RNases found in Arabidopsis, and preliminary evidence shows that it may be involved in phosphate recycling and localized to an intracellular compartment. Using rns2 knock-out mutant plants, I showed that RNS2 has an effect on flowering time during times of phosphate starvation. I also showed that RNS2 is localized to the ER, more specifically, to ER-bodies. To ...


Production Of 3-Hydroxypropionate From Biomass, Netra Rajguru Agarkar Jan 2007

Production Of 3-Hydroxypropionate From Biomass, Netra Rajguru Agarkar

Retrospective Theses and Dissertations

Biorenewable technology is a developing field of science that researches alternative sources for petroleum products like fuels, plastics, paints, etc. The DoE biomass program has identified the 12 top chemicals that can be produced from biomass and further processed to replace many petroleum products; 3-hydroxypropionate (3HP) is a top 12 chemical that can be used to produce plastics, paints, tires, and other consumer products. This thesis describes the attempts to produce 3HP from glucose in E. coli and from syngas in R. rubrum. Malonyl CoA reductase from C. aurantiacus and R. castenholzii was cloned and expressed in E. coli and ...


Methanobactin And The Membrane-Associated Methane Monooxygenase In Methanotrophy: A Tale Of Two Proteins, Dongwon Choi Jan 2007

Methanobactin And The Membrane-Associated Methane Monooxygenase In Methanotrophy: A Tale Of Two Proteins, Dongwon Choi

Retrospective Theses and Dissertations

No abstract provided.


Characterization Of Three Spindle Matrix Proteins, Megator, East And Asator, In Drosophila, Hongying Qi Jan 2007

Characterization Of Three Spindle Matrix Proteins, Megator, East And Asator, In Drosophila, Hongying Qi

Retrospective Theses and Dissertations

Two previously identified proteins Skeletor and Chromator provide the molecular evidence for the existence of a spindle matrix complex in Drosophila. The spindle matrix has been proposed to be a macromolecular complex, which plays a role in organization and stabilization of the mitotic spindle as well as in providing structural support for counterbalancing force production.;In this dissertation, I present the identification and characterization of three more putative spindle matrix candidate proteins, Megator, EAST and Asator. Megator, an ortholog of the mammalian TPR protein, contains a large coiled-coil domain at its NH2-terminus and an acidic, non-structural COOH-terminus. Immunohistochemistry studies show ...


Biophysical Characterization Of The 117 Amino Acids Long N-Terminal Segment Of D-Raf (Isoform A), Oren Tchaicheeyan Jan 2007

Biophysical Characterization Of The 117 Amino Acids Long N-Terminal Segment Of D-Raf (Isoform A), Oren Tchaicheeyan

Retrospective Theses and Dissertations

Drosophila Raf (D-Raf) is an orthologue of Human B-Raf. D-Raf may have two splice variants, a short (Isoform A, I-A) and a long one (Isoform B, I-B). Protocols for over-expression (E. Coli) and purification of 117 amino acids (aa) N-terminal segment (NTS) of D-Raf protein (I-A), both WT and Mutant (T60E) were established. These protein constructs were also biophysically characterized. Thrombin cleavage conditions were optimized, as monitored by 5 amino acid NTS sequencing, mass spectroscopy and SDS-PAGE electrophoresis. An attempt to purify the 160 aa D-Raf (I-B) NTS failed. NMR characterization in a 700 MHz magnet employing three experiments (HSQC ...


Allosteric Regulation Of Mammalian And Bacterial Fructose-1,6-Bisphosphatases, Justin Keith Hines Jan 2007

Allosteric Regulation Of Mammalian And Bacterial Fructose-1,6-Bisphosphatases, Justin Keith Hines

Retrospective Theses and Dissertations

Fructose-1,6-bisphosphatase (FBPase) is a critical regulatory enzyme in gluconeogenesis. In mammals, the enzyme is subject to metabolic regulation; the hormones insulin and glucagon reciprocally regulate the enzyme in the liver through the controlled production and degradation of the inhibitor fructose-2,6-bisphosphate (Fru-2,6-P2) and hence control hepatic glucose production. FBPase is nearly ubiquitous in living organisms and in the bacterium Escherichia coli , the homologous enzyme is imperative for growth on gluconeogenic substrates. How does this organism regulate FBPase without the use of exogenous hormones or Fru-2,6-P2? Investigations reported herein describe high resolution crystal structures of the enzyme from ...


Determinants Of Substrate Recognition And Mechanisms Of Subunit Exchange, Yang Zhou Jan 2007

Determinants Of Substrate Recognition And Mechanisms Of Subunit Exchange, Yang Zhou

Retrospective Theses and Dissertations

Adenylosuccinate synthetase (ADSS) offers a good opportunity to measure the effect of non-bonded interactions. Both IMP and 2'-deoxy-IMP are good substrates supporting identical binding affinities and maximal velocities; however, L-aspartate exhibits significant differences in binding affinities depending on which nucleotide is used as a substrate. Crystal structures of both complexes were identical except for the absence of the 2'-OH group in the dexoy-nucleotide complex. The decrease in energy (∼2 kcal/mole) in non-bonded interactions due to the loss of one atom explains the nearly 40-fold difference in the binding affinity of L-aspartate and an analog of L-aspartate called ...


Investigation Of Allosteric Regulation Of Porcine Fructose-1,6-Bisphosphatase, Jian Lu Jan 2007

Investigation Of Allosteric Regulation Of Porcine Fructose-1,6-Bisphosphatase, Jian Lu

Retrospective Theses and Dissertations

Porcine Fructose-1,6-bisphosphatase is a homotetramer with four identical subunits. It plays a central role in gluconeogenesis and is tightly regulated by metabolites fructose 2,6-bisphosphate (F26P2) and AMP. Loop 52-72, loop 182-194 and residues 7-11 of the N-terminal segment play important role in the mechanism of catalysis and allosteric inhibition by AMP by retaining a structural hydrophobic region. Disruption of the hydrophobic region will evidently affect the catalysis and regulation of FBPase. Mutation of Thr53 reduced the catalysis activity of FBPase, mutation of Thr194 eliminated inhibition of AMP due to its low affinity to the enzyme, while double mutation ...


Structural And Functional Studies On Membrane Fusion Proteins-Snares, Yibin Xu Jan 2007

Structural And Functional Studies On Membrane Fusion Proteins-Snares, Yibin Xu

Retrospective Theses and Dissertations

Membrane fusion is a ubiquitous and essential intracellular function. Membrane fusion is mediated by a protein superfamily known as N-ethymaleimide-sensitive fusion protein attachment protein receptors (SNAREs). From plant to mammal, there is a remarkable sequence similarity among the SNARE-family related to the conserved function of this family. The function of SNAREs has been proposed to be the key to membrane fusion as minimal fusion machinery. Depending on their distinct membrane distribution, SNAREs were classified into v (vesicle)-SNAREs and t (target)-SNAREs. It is widely believed that SNARE complex assembly by v- and t-SNAREs on opposite membranes offers the ultimate ...


Structural Study Of Yeast Snare Proteins On Membrane By Epr, Yinghui Zhang Jan 2007

Structural Study Of Yeast Snare Proteins On Membrane By Epr, Yinghui Zhang

Retrospective Theses and Dissertations

Membrane fusion is a basic biophysical process involved in cell's physiological activities. The value of studying the exocytosis of mammalian cells and yeast cells lies in that a highly conserved SNARE (Soluble N-ethylmaleimide-sensitive factor attachment protein receptor) protein super family mediates this process and is proposed to be the core protein machinery for the membrane. The SNARE proteins located on plasma membrane and vesicle membrane are assembled together to form a trans-SNARE bridging two separate membranes and initiating membrane.;My research centers on how SNARE assembly involved in yeast cells' exocytosis promotes membrane fusion between yeast vesicle and plasma ...


Kinetic Studies Of Escherichia Coli And Human Saicar Synthetase, Daniel Joseph Binkowski Jan 2007

Kinetic Studies Of Escherichia Coli And Human Saicar Synthetase, Daniel Joseph Binkowski

Retrospective Theses and Dissertations

Phosphoribosyl-aminoimidazole-succinocarboxamide synthetase (SAICAR synthetase) catalyzes the eighth step of de novo IMP biosynthesis in bacteria, and the seventh step in humans. The SAICAR synthetase reaction is analogous to that of adenylosuccinate synthetase (first committed step of AMP biosynthesis), using ATP to ligate 5-amino-imidazole-4-carboxy ribonucleotide and L-aspartate to produce 5-aminoimidazole-4(N-succinylcarboxamide) ribonucleotide (SAICAR). SAICAR synthetase and other enzymes of purine nucleotide biosynthesis are targets of natural products that impair cell growth. Prior to these studies, the kinetic mechanism of any SAICAR synthetase was unknown. Herein are reported the kinetic mechanisms of bacterial and human SAICAR synthetase activities, the structure of the ...


Crystallographic Analysis Of Parasponia Andersonii Hemoglobin, Andrea Savage Jan 2007

Crystallographic Analysis Of Parasponia Andersonii Hemoglobin, Andrea Savage

Retrospective Theses and Dissertations

The x-ray crystal structure of ferric Parasponia andersonii hemoglobin has been determined to 2.3 A resolution by molecular replacement with Rice hemoglobin 1 (rHb1). By means of absorbance spectroscopy, electrochemistry and x-ray crystallography, it has been revealed that depending on the oxidation state, the heme iron can exsist in either the pentacoordinate or hexacoordinate form. Pentacoordination of the heme iron is found primarily in the deoxy ferrous (2+) form, whereas the ferric (3+) form is exhibited predominantly in bis-histidine coordination. This is unique among known forms of hemoglobins. Furthermore, the crystallographic data has provided evidence which supports previous works ...


Characterization Of The Starch Branching Enzymes In Arabidopsis Thaliana, Qinglei Gan Jan 2007

Characterization Of The Starch Branching Enzymes In Arabidopsis Thaliana, Qinglei Gan

Retrospective Theses and Dissertations

The roles of three starch branching enzyme isoforms, BE1, BE2 and BE3, in Arabidopsis leaf starch metabolism were investigated by AtBE mutant analysis. We also determined the expression patterns of BEs in Arabidopsis by GUS staining and microarray experiments. The possible correlation of BE gene expression with other genes in the Arabidopsis genome was analyzed to identify putative genes that are functionally correlated with BEs. Motifs in the promoters of the Arabidopsis BE genes were identified to better understand their expression differences. BE2 and BE3 expression levels during a diurnal cycle were evaluated with isoform specific antibodies. Meantime, the in ...


Role Of Leu[Superscript B 10] In Cytoglobin And Comparison Of Direct And Indirect Measurements Of Hexacoordinate Hemoglobin Ligand Affinity, Jordan Richard Witmer Jan 2007

Role Of Leu[Superscript B 10] In Cytoglobin And Comparison Of Direct And Indirect Measurements Of Hexacoordinate Hemoglobin Ligand Affinity, Jordan Richard Witmer

Retrospective Theses and Dissertations

Hexacoordinate hemoglobins (HxHbs) have recently been discovered and have been found in nearly all living organisms. In humans, hxHbs are represented by neuroglobin (Ngb) and cytoglobin (Cgb). While Ngb is principally found in the brain, Cgb is ubiquitously expressed in all tissues. Ligand binding in hexacoordinate hemoglobins is complex due to the reversible coordination of the heme iron by the distal histidine. Hexacoordination regulates ligand binding and is regulated by specific residues in the heme pocket including the B10 position. Therefore, we investigated the influence of LeuB10 on hexacoordination and ligand binding in Cgb. Ligand binding was analyzed for recombinant ...


Biochemical Studies Of Mms6--A Biomineralization Protein, Lijun Wang Jan 2007

Biochemical Studies Of Mms6--A Biomineralization Protein, Lijun Wang

Retrospective Theses and Dissertations

Magnetotactic bacteria are aquatic prokaryotes that can move under the direction of a local geomagnetic field. The magnetosomes found in magnetotactic bacteria are arranged in a single chain or chains, which is the physics basis of magnetotaxis trait. It is believed that with the trait of magnetotaxis, magnetotactic bacteria can simplify their search for the optimal oxygen environments.;Mms6 is a magnetosome membrane-associated protein in Magnetospirillum magneticum strain AMB-1. It has 59 amino acids with a hydrophobic N-terminal and a hydrophilic C-terminal region. Magnetite nanoparticles with similar size and shape to bacterial magnetite can be synthesized in vitro in the ...


The Role Of Tyr61 In Synechocystis Hemoglobin In Regulating E-Helix Movement, Anand Venugopal Jan 2007

The Role Of Tyr61 In Synechocystis Hemoglobin In Regulating E-Helix Movement, Anand Venugopal

Retrospective Theses and Dissertations

Hexacoordinate hemoglobins have been discovered that have yet to have a functional role confidently assigned to them. The eubacteria synechocystis sp. PCC 6803 has been found to have a truncated hemoglobin which is hexacoordinated. The structures of the hexacoordinated and cyanide bound forms of synechocystis hemoglobin have been solved, and a large shift in the E-helix has been seen upon ligand binding. Also, the loop connecting the E and F helix contains residues Tyr61 and Asp62 that are involved in hydrogen bonding networks in the cyanide-bound and hexacoordinate states respectively. A mutation was made to the Y61 residue to make ...


Biophysical And Biochemical Characterization Of Hexacoordinate Hemoglobins, Benoit Joel Smagghe Jan 2007

Biophysical And Biochemical Characterization Of Hexacoordinate Hemoglobins, Benoit Joel Smagghe

Retrospective Theses and Dissertations

Within the hemoglobin (Hb) superfamily, an unusual group called hexacoordinate hemoglobin (hxHbs) whose members are found in all living organisms is the subject of exciting studies since their discovery. They are named "hexacoordinate" because of reversible coordination of the sixth coordination site of the heme iron by a histidine side chain located in the heme distal pocket. Interestingly, hxHbs share high degree of similarity in their primary sequence and a common globin fold indicating that they might share a common role and that endogenous coordination might be an important part of their not yet characterized physiological function. The characterization of ...