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Molecular Biology

2008

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Articles 1 - 30 of 149

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

Molecular Characterisation Of Canine Nonsteroidal Anti-Inflammatory Drug-Activated Gene (Nag-1), K Yamaguchi, Nichelle Whitlock, Jason Liggett, Alfred Legendre, Michael Fry, Seung Baek Dec 2010

Molecular Characterisation Of Canine Nonsteroidal Anti-Inflammatory Drug-Activated Gene (Nag-1), K Yamaguchi, Nichelle Whitlock, Jason Liggett, Alfred Legendre, Michael Fry, Seung Baek

Alfred M Legendre DVM, MS, DACVIM

Nonsteroidal anti-inflammatory drug (NSAID)-activated gene (NAG-1), a divergent member of the transforming growth factor beta superfamily, was previously identified as a gene induced by several anti-tumorigenic compounds, including NSAIDs and peroxisome proliferator-activated receptor gamma (PPARgamma) ligands in humans. In this study, canine NAG-1 was characterised from a canine genomic database. Gene induction by some NSAIDs and PPARgamma ligands was demonstrated in canine osteosarcoma cell lines. Phylogenetic analysis indicates that canine NAG-1 is more homologous with the corresponding mouse and rat genes than with human NAG-1. Expression of canine NAG-1 was increased by treatment with piroxicam and SC-560 (NSAIDs) and ...


Multiple Mechanisms Are Involved In 6-Gingerol-Induced Cell Growth Arrest And Apoptosis In Human Colorectal Cancer Cells, Seong Lee, Maria Cekanova, Seung Baek Oct 2010

Multiple Mechanisms Are Involved In 6-Gingerol-Induced Cell Growth Arrest And Apoptosis In Human Colorectal Cancer Cells, Seong Lee, Maria Cekanova, Seung Baek

Maria Cekanova MS, RNDr, PhD

6-Gingerol, a natural product of ginger, has been known to possess anti-tumorigenic and pro-apoptotic activities. However, the mechanisms by which it prevents cancer are not well understood in human colorectal cancer. Cyclin D1 is a proto-oncogene that is overexpressed in many cancers and plays a role in cell proliferation through activation by beta-catenin signaling. Nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1) is a cytokine associated with pro-apoptotic and anti-tumorigenic properties. In the present study, we examined whether 6-gingerol influences cyclin D1 and NAG-1 expression and determined the mechanisms by which 6-gingerol affects the growth of human colorectal cancer cells in ...


Peroxisome Proliferator-Activated Receptor Ligand Mcc-555 Suppresses Intestinal Polyps In Apcmin/+ Mice Via Extracellular Signal-Regulated Kinase And Peroxisome Proliferator-Activated Receptor-Dependent Pathways, K Yamaguchi, Maria Cekanova, Michael Mcentee, J Yoon, S Fischer, I Renes, I Van Seungnigen, Seung Baek Oct 2010

Peroxisome Proliferator-Activated Receptor Ligand Mcc-555 Suppresses Intestinal Polyps In Apcmin/+ Mice Via Extracellular Signal-Regulated Kinase And Peroxisome Proliferator-Activated Receptor-Dependent Pathways, K Yamaguchi, Maria Cekanova, Michael Mcentee, J Yoon, S Fischer, I Renes, I Van Seungnigen, Seung Baek

Maria Cekanova MS, RNDr, PhD

A large body of studies has suggested that peroxisome proliferator-activated receptor gamma (PPARgamma) ligands, such as thiazolidinedione, are potent candidates for chemopreventive agents. MCC-555 is a PPARgamma/alpha dual agonist and has been shown previously to induce apoptosis in vitro; however, the molecular mechanisms by which MCC-555 affects antitumorigenesis in vivo are poorly understood. In this study, we explored the antitumorigenic effects of MCC-555 both in cell culture and in Apc-deficient mice, an animal model for human familial adenomatous polyposis. MCC-555 increased MUC2 expression in colorectal and lung cancer cells, and treatment with the PPARgamma antagonist GW9662 revealed that MUC2 ...


Structure And Dynamics Of Metalloproteins In Live Cells, Jeremy D. Cook, James E. Penner-Hahn, Timothy L. Stemmler Dec 2008

Structure And Dynamics Of Metalloproteins In Live Cells, Jeremy D. Cook, James E. Penner-Hahn, Timothy L. Stemmler

Biochemistry and Molecular Biology Faculty Publications

X-ray absorption spectroscopy (XAS) has emerged as one of the premier tools for investigating the structure and dynamic properties of metals in cells and in metal containing biomolecules. Utilizing the high flux and broad energy range of X-rays supplied by synchrotron light sources, one can selectively excite core electronic transitions in each metal. Spectroscopic signals from these electronic transitions can be used to dissect the chemical architecture of metals in cells, in cellular components and in biomolecules at varying degrees of structural resolution. With the development of ever-brighter X-ray sources, X-ray methods have grown into applications that can be utilized ...


A Genetically Encoded Fluorescent Sensor Of Erk Activity, Christopher D. Harvey, Anka G. Ehrhardt, Cristina Arrigo Cellurale, Haining Zhong, Ryohei Yasuda, Roger J. Davis, Karel Svoboda Dec 2008

A Genetically Encoded Fluorescent Sensor Of Erk Activity, Christopher D. Harvey, Anka G. Ehrhardt, Cristina Arrigo Cellurale, Haining Zhong, Ryohei Yasuda, Roger J. Davis, Karel Svoboda

Davis Lab Publications

The activity of the ERK has complex spatial and temporal dynamics that are important for the specificity of downstream effects. However, current biochemical techniques do not allow for the measurement of ERK signaling with fine spatiotemporal resolution. We developed a genetically encoded, FRET-based sensor of ERK activity (the extracellular signal-regulated kinase activity reporter, EKAR), optimized for signal-to-noise ratio and fluorescence lifetime imaging. EKAR selectively and reversibly reported ERK activation in HEK293 cells after epidermal growth factor stimulation. EKAR signals were correlated with ERK phosphorylation, required ERK activity, and did not report the activities of JNK or p38. EKAR reported ERK ...


Comparison Of Electrically Mediated And Liposome-Complexed Plasmid Dna Delivery To The Skin, Loree C. Heller, Mark J. Jaroszeski, Domenico Coppola, Richard Heller Dec 2008

Comparison Of Electrically Mediated And Liposome-Complexed Plasmid Dna Delivery To The Skin, Loree C. Heller, Mark J. Jaroszeski, Domenico Coppola, Richard Heller

Bioelectrics Publications

BACKGROUND: Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery.

METHODS: Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model.

RESULTS: Experiments confirm ...


Ese-1/Egr-1 Pathway Plays A Role In Tolfenamic Acid-Induced Apoptosis In Colorectal Cancer Cells, Seong Lee, J Bahn, C Choi, Nichelle Whitlock, A English, S Safe, Seung Baek Nov 2008

Ese-1/Egr-1 Pathway Plays A Role In Tolfenamic Acid-Induced Apoptosis In Colorectal Cancer Cells, Seong Lee, J Bahn, C Choi, Nichelle Whitlock, A English, S Safe, Seung Baek

Seung J Baek

Nonsteroidal anti-inflammatory drugs (NSAIDs) are known to prevent colorectal tumorigenesis. Although antitumor effects of NSAIDs are mainly due to inhibition of cyclooxygenase activity, there is increasing evidence that cyclooxygenase-independent mechanisms may also play an important role. The early growth response-1 (EGR-1) gene is a member of the immediate-early gene family and has been identified as a tumor suppressor gene. Tolfenamic acid is a NSAID that exhibits anticancer activity in a pancreatic cancer model. In the present study, we investigated the anticancer activity of tolfenamic acid in human colorectal cancer cells. Tolfenamic acid treatment inhibited cell growth and induced apoptosis as ...


Inhibition Of Yeast Telomerase Action By The Telomeric Ssdna-Binding Protein, Cdc13p, David C. Zappulla, Jennifer N. Roberts, Karen J. Goodrich, Thomas R. Cech, Deborah S. Wuttke Nov 2008

Inhibition Of Yeast Telomerase Action By The Telomeric Ssdna-Binding Protein, Cdc13p, David C. Zappulla, Jennifer N. Roberts, Karen J. Goodrich, Thomas R. Cech, Deborah S. Wuttke

Chemistry & Biochemistry Faculty Contributions (1986-2018)

Appropriate control of the chromosome end-replicating enzyme telomerase is crucial for maintaining telomere length and genomic stability. The essential telomeric DNA-binding protein Cdc13p both positively and negatively regulates telomere length in budding yeast. Here we test the effect of purified Cdc13p on telomerase action in vitro. We show that the full-length protein and its DNA-binding domain (DBD) inhibit primer extension by telomerase. This inhibition occurs by competitive blocking of telomerase access to DNA. To further understand the requirements for productive telomerase 3′-end access when Cdc13p or the DBD is bound to a telomerase substrate, we constrained protein binding at ...


The Cooh-Terminal Domain Of The Jil-1 Histone H3s10 Kinase Interacts With Histone H3 And Is Required For Correct Targeting To Chromatin, Xiaomin Bao, Weili Cai, Huai Deng, Weiguo Zhang, Robert Krencik, Jack Girton, Jorgen Johansen, Kristen M. Johansen Nov 2008

The Cooh-Terminal Domain Of The Jil-1 Histone H3s10 Kinase Interacts With Histone H3 And Is Required For Correct Targeting To Chromatin, Xiaomin Bao, Weili Cai, Huai Deng, Weiguo Zhang, Robert Krencik, Jack Girton, Jorgen Johansen, Kristen M. Johansen

Biochemistry, Biophysics and Molecular Biology Publications

The JIL-1 histone H3S10 kinase in Drosophila localizes specifically to euchromatic interband regions of polytene chromosomes and is enriched 2-fold on the male X chromosome. JIL-1 can be divided into four main domains including an NH2-terminal domain, two separate kinase domains, and a COOH-terminal domain. Our results demonstrate that the COOH-terminal domain of JIL-1 is necessary and sufficient for correct chromosome targeting to autosomes but that both COOH- and NH2-terminal sequences are necessary for enrichment on the male X chromosome. We furthermore show that a small 53-amino acid region within the COOH-terminal domain can interact with ...


Orientational Distributions Of Contact Clusters In Proteins Closely Resemble Those Of An Icosahedron, Yaping Feng, Robert L. Jernigan, Andrzej Kloczkowski Nov 2008

Orientational Distributions Of Contact Clusters In Proteins Closely Resemble Those Of An Icosahedron, Yaping Feng, Robert L. Jernigan, Andrzej Kloczkowski

Biochemistry, Biophysics and Molecular Biology Publications

The orientational geometry of residue packing in proteins was studied in the past by superimposing clusters of neighboring residues with several simple lattices.1,2 In this work, instead of a lattice we use the regular polyhedron, the icosahedron, as the model to describe the orientational distribution of contacts in clusters derived from a high-resolution protein dataset (522 protein structures with high resolution < 1.5Å). We find that the order parameter (orientation function) measuring the angular overlap of directions in coordination clusters with directions of the icosahedron is 0.91, which is a significant improvement in comparison with the value 0.82 for the order parameter with the face-centered cubic (fcc) lattice. Close packing tendencies and patterns of residue packing in proteins is considered in detail and a theoretical description of these packing regularities is proposed.


Reexamination Of The Role Of The Amino Terminus Of Seca In Promoting Its Dimerization And Functional State, Don Oliver Nov 2008

Reexamination Of The Role Of The Amino Terminus Of Seca In Promoting Its Dimerization And Functional State, Don Oliver

Division III Faculty Publications

No abstract provided.


Web-Based Arabidopsis Functional And Structural Genomics Resources, Yan Lu, Robert Last Oct 2008

Web-Based Arabidopsis Functional And Structural Genomics Resources, Yan Lu, Robert Last

Yan Lu

No abstract provided.


Conversion Of Red Fluorescent Protein Into A Bright Blue Probe, Oksana M. Subach, Illia S. Gundorov, Masami Yoshimura, Fedor V. Subach, Jinghang Zhang, David Grunwald, Ekaterina A. Souslova, Dmitriy M. Chudakov, Vladislav V. Verkhusha Oct 2008

Conversion Of Red Fluorescent Protein Into A Bright Blue Probe, Oksana M. Subach, Illia S. Gundorov, Masami Yoshimura, Fedor V. Subach, Jinghang Zhang, David Grunwald, Ekaterina A. Souslova, Dmitriy M. Chudakov, Vladislav V. Verkhusha

Biochemistry and Molecular Pharmacology Publications and Presentations

We used a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a rational design strategy to develop blue fluorescent proteins with a tyrosine-based chromophore. The strategy was applied to red fluorescent proteins of the different genetic backgrounds, such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which all were converted into blue probes. Further improvement of the blue variant of TagRFP by random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by the substantially higher brightness, the faster chromophore maturation, and the higher pH stability than blue fluorescent proteins ...


Dynamics Of Apomyoglobin In The Α-To-Β Transition And Of Partially Unfolded Aggregated Protein, E. Fabiani, A. M. Stadler, D. Madern, M. M. Koza, M. Tehei, M. Hirai, G. Zaccai Oct 2008

Dynamics Of Apomyoglobin In The Α-To-Β Transition And Of Partially Unfolded Aggregated Protein, E. Fabiani, A. M. Stadler, D. Madern, M. M. Koza, M. Tehei, M. Hirai, G. Zaccai

Faculty of Science - Papers (Archive)

Changes of molecular dynamics in the α-to-β transition associated with amyloid fibril formation were explored on apo-myoglobin (ApoMb) as a model system. Circular dichroism, neutron and X-ray scattering experiments were performed as a function of temperature on the protein, at different solvent conditions. A significant change in molecular dynamics was observed at the α-to-β transition at about 55 ˚C, indicating a more resilient high temperature β structure phase. A similar effect at approximately the same temperature was observed in holo-myoglobin, associated with partial unfolding and protein aggregation. A study in a wide temperature range between 20 K and 360 K ...


Curriculum Vitae, Robert P. Lane Oct 2008

Curriculum Vitae, Robert P. Lane

Robert P. Lane

No abstract provided.


Autonomy And Robustness Of Translocation Through The Nuclear Pore Complex: A Single-Molecule Study, Thomas Dange, David Grunwald, Antje Grunwald, Reiner Peters, Ulrich Kubitscheck Oct 2008

Autonomy And Robustness Of Translocation Through The Nuclear Pore Complex: A Single-Molecule Study, Thomas Dange, David Grunwald, Antje Grunwald, Reiner Peters, Ulrich Kubitscheck

Biochemistry and Molecular Pharmacology Publications and Presentations

All molecular traffic between nucleus and cytoplasm occurs via the nuclear pore complex (NPC) within the nuclear envelope. In this study we analyzed the interactions of the nuclear transport receptors kapalpha2, kapbeta1, kapbeta1DeltaN44, and kapbeta2, and the model transport substrate, BSA-NLS, with NPCs to determine binding sites and kinetics using single-molecule microscopy in living cells. Recombinant transport receptors and BSA-NLS were fluorescently labeled by AlexaFluor 488, and microinjected into the cytoplasm of living HeLa cells expressing POM121-GFP as a nuclear pore marker. After bleaching the dominant GFP fluorescence the interactions of the microinjected molecules could be studied using video microscopy ...


Transient High Glucose Causes Persistent Epigenetic Changes And Altered Gene Expression During Subsequent Normoglycemia, Assam El-Osta, Daniella Brasacchio, Dachun Yao, Alessandro Pocai, Peter L. Jones, Robert G. Roeder, Mark E. Cooper, Michael Brownlee Sep 2008

Transient High Glucose Causes Persistent Epigenetic Changes And Altered Gene Expression During Subsequent Normoglycemia, Assam El-Osta, Daniella Brasacchio, Dachun Yao, Alessandro Pocai, Peter L. Jones, Robert G. Roeder, Mark E. Cooper, Michael Brownlee

Peter Jones Lab Publications

The current goal of diabetes therapy is to reduce time-averaged mean levels of glycemia, measured as HbA1c, to prevent diabetic complications. However, HbA1c only explains <25% of the variation in risk of developing complications. Because HbA1c does not correlate with glycemic variability when adjusted for mean blood glucose, we hypothesized that transient spikes of hyperglycemia may be an HbA1c-independent risk factor for diabetic complications. We show that transient hyperglycemia induces long-lasting activating epigenetic changes in the promoter of the nuclear factor kappaB (NF-kappaB) subunit p65 in aortic endothelial cells both in vitro and in nondiabetic mice, which cause increased p65 gene expression. Both the epigenetic changes and the gene expression changes persist for at least 6 d of subsequent normal glycemia, as do NF-kappaB-induced increases in monocyte chemoattractant protein 1 and vascular cell adhesion molecule 1 expression. Hyperglycemia-induced epigenetic changes and increased p65 expression are prevented by reducing mitochondrial superoxide production or superoxide-induced alpha-oxoaldehydes. These results highlight the dramatic and long-lasting effects that short-term hyperglycemic spikes can have on vascular cells and suggest that transient spikes of hyperglycemia may be an HbA1c-independent risk factor for diabetic complications.


Toward Predicting Self-Splicing And Protein-Facilitated Splicing Of Group I Introns, Quentin Vicens, Paul J. Paukstelis, Eric Westhof, Alan M. Lambowitz, Thomas R. Cech Sep 2008

Toward Predicting Self-Splicing And Protein-Facilitated Splicing Of Group I Introns, Quentin Vicens, Paul J. Paukstelis, Eric Westhof, Alan M. Lambowitz, Thomas R. Cech

Chemistry & Biochemistry Faculty Contributions (1986-2018)

In the current era of massive discoveries of noncoding RNAs within genomes, being able to infer a function from a nucleotide sequence is of paramount interest. Although studies of individual group I introns have identified self-splicing and nonself-splicing examples, there is no overall understanding of the prevalence of self-splicing or the factors that determine it among the >2300 group I introns sequenced to date. Here, the self-splicing activities of 12 group I introns from various organisms were assayed under six reaction conditions that had been shown previously to promote RNA catalysis for different RNAs. Besides revealing that assessing self-splicing under ...


Peroxisome Proliferator-Activated Receptor Ligand Mcc-555 Suppresses Intestinal Polyps In Apcmin/+ Mice Via Extracellular Signal-Regulated Kinase And Peroxisome Proliferator-Activated Receptor-Dependent Pathways, K Yamaguchi, Maria Cekanova, Michael Mcentee, J Yoon, S Fischer, I Renes, I Van Seungnigen, Seung Baek Aug 2008

Peroxisome Proliferator-Activated Receptor Ligand Mcc-555 Suppresses Intestinal Polyps In Apcmin/+ Mice Via Extracellular Signal-Regulated Kinase And Peroxisome Proliferator-Activated Receptor-Dependent Pathways, K Yamaguchi, Maria Cekanova, Michael Mcentee, J Yoon, S Fischer, I Renes, I Van Seungnigen, Seung Baek

Seung J Baek

A large body of studies has suggested that peroxisome proliferator-activated receptor gamma (PPARgamma) ligands, such as thiazolidinedione, are potent candidates for chemopreventive agents. MCC-555 is a PPARgamma/alpha dual agonist and has been shown previously to induce apoptosis in vitro; however, the molecular mechanisms by which MCC-555 affects antitumorigenesis in vivo are poorly understood. In this study, we explored the antitumorigenic effects of MCC-555 both in cell culture and in Apc-deficient mice, an animal model for human familial adenomatous polyposis. MCC-555 increased MUC2 expression in colorectal and lung cancer cells, and treatment with the PPARgamma antagonist GW9662 revealed that MUC2 ...


The Role Of Gtp Binding And Hydrolysis At The Attoc159 Preprotein Receptor During Protein Import Into Chloroplasts, Danny Schnell, F. Wang, B. Agne, F. Kessler Aug 2008

The Role Of Gtp Binding And Hydrolysis At The Attoc159 Preprotein Receptor During Protein Import Into Chloroplasts, Danny Schnell, F. Wang, B. Agne, F. Kessler

Danny Schnell

The majority of nucleus-encoded chloroplast proteins are targeted to the organelle by direct binding to two membrane-bound GTPase receptors, Toc34 and Toc159. The GTPase activities of the receptors are implicated in two key import activities, preprotein binding and driving membrane translocation, but their precise functions have not been defined. We use a combination of in vivo and in vitro approaches to study the role of the Toc159 receptor in the import reaction. We show that atToc159-A864R, a receptor with reduced GTPase activity, can fully complement a lethal insertion mutation in the ATTOC159 gene. Surprisingly, the atToc159-A864R receptor increases the rate ...


Characterization Of Heat Shock Protein A12b As A Novel Angiogenesis Regulator., Rebecca J. Steagall Aug 2008

Characterization Of Heat Shock Protein A12b As A Novel Angiogenesis Regulator., Rebecca J. Steagall

Electronic Theses and Dissertations

Previously, we cloned Heat shock protein A12B (HspA12B), the newest member of a recently defined subfamily of proteins distantly related to the Hsp70 family that are enriched in atherosclerotic lesions. We have found that HspA12B is predominantly expressed in vascular endothelium, and that it is involved in angiogenesis which we probed by in vitro angiogenesis assays (Matrigel), migration assays and Directed In Vivo Angiogenesis Assay (DIVAA). Hsp70s are molecular chaperones that are inducible by stress and have been found to be anti-apoptotic (Li et al. 2000; Nylandsted et al. 2000; Garrido et al. 2001). Because of its homology to Hsp70 ...


Detection Of Biomarkers By Mrs Requires Error Analysis, Jeffrey C. Hoch, Mark W. Maciejewski, Michael R. Gryk Aug 2008

Detection Of Biomarkers By Mrs Requires Error Analysis, Jeffrey C. Hoch, Mark W. Maciejewski, Michael R. Gryk

UCHC Articles - Research

Using MRS imaging and singular value decomposition (SVD), Manganas et al. (Science 318, 980,2007) reported a nuclear magnetic resonance diagnostic for neural progenitor cells. Characteristics of MRS experiments are known to be problematic for SVD-based methods. Absent detection using alternate methods of spectrum analysis or controls to quantify the false discovery rate, little significance can be attached to their result.


Bacterial Expression And Purification Of Interleukin-2 Tyrosine Kinase: Single Step Separation Of The Chaperonin Impurity, Raji E. Joseph, Amy H. Andreotti Aug 2008

Bacterial Expression And Purification Of Interleukin-2 Tyrosine Kinase: Single Step Separation Of The Chaperonin Impurity, Raji E. Joseph, Amy H. Andreotti

Biochemistry, Biophysics and Molecular Biology Publications

Biochemical and biophysical characterization of kinases requires large quantities of purified protein. Here we report the bacterial expression and purification of active Itk kinase domain (a Tec family kinase) using ArcticExpress cells that co-express the chaperonin system Cpn60/10 from Oleispira antarctica. We describe a simple one step MgCl2/ATP/KCl incubation procedure to remove the copurifying chaperonin impurity. Chaperonin co-purification is a common problem encountered during protein purification and the simple incubation step described here completely overcomes this problem. The approach targets the chaperonin system rather than the protein of interest and is therefore widely applicable to other protein ...


Heteromeric Interactions Among Ethylene Receptors Mediate Signaling In Arabidopsis, Brad Binder, Zhiyong Gao, Chi-Kuang Wen, Yi-Feng Chen, Jianhong Chang, Yi-Hsuan Chiang, Robert Kerris, Caren Chang, G. Schaller Jul 2008

Heteromeric Interactions Among Ethylene Receptors Mediate Signaling In Arabidopsis, Brad Binder, Zhiyong Gao, Chi-Kuang Wen, Yi-Feng Chen, Jianhong Chang, Yi-Hsuan Chiang, Robert Kerris, Caren Chang, G. Schaller

Brad M. Binder

The gaseous hormone ethylene is perceived in Arabidopsis by a five member receptor family that consists of the subfamily 1 receptors ETR1 and ERS1 and the subfamily 2 receptors ETR2, ERS2, and EIN4. Previous work has demonstrated that the basic functional unit for the ethylene receptor, ETR1, is a disulfide-linked homodimer. We demonstrate here that ethylene receptors isolated from Arabidopsis also interact with each other through noncovalent interactions. Evidence that ETR1 associates with other ethylene receptors was obtained by co-purification of ETR1 with tagged versions of ERS1, ETR2, ERS2, and EIN4 from Arabidopsis membrane extracts. ETR1 preferentially associated with the ...


Epigallocatechin-3-Gallate Inhibits Interleukin-1beta-Induced Muc5ac Gene Expression And Muc5ac Secretion In Normal Human Nasal Epithelial Cells, H Kim, S Park, S Park, U Moon, B Lee, S Yoon, J Lee, Seung Baek, J Yoon Jul 2008

Epigallocatechin-3-Gallate Inhibits Interleukin-1beta-Induced Muc5ac Gene Expression And Muc5ac Secretion In Normal Human Nasal Epithelial Cells, H Kim, S Park, S Park, U Moon, B Lee, S Yoon, J Lee, Seung Baek, J Yoon

Seung J Baek

It has been reported that the proinflammatory cytokine interleukin-1beta (IL-1beta) induces mucus hypersecretion in normal human nasal epithelial (NHNE) cells and that the MAP kinase pathway may be an important signal pathway in IL-1beta-induced MUC5AC gene expression. Green tea (Camellia sinensis) polyphenols are potent anti-inflammatory agents and have been shown to inhibit inflammation in tumor cell lines and cultured respiratory epithelial cells. In this study, we examined the effect of (-)-epigallocatechin-3-gallate (EGCG), a green tea polyphenol, on IL-1beta-induced MUC5AC gene expression and secretion in NHNE cells. After cells had been treated with IL-1beta (10 ng/ml) and pretreated with EGCG ...


Dendrimer Supramolecular Assembly For Gene Delivery, Karthikeyan Pasupathy Jul 2008

Dendrimer Supramolecular Assembly For Gene Delivery, Karthikeyan Pasupathy

All Theses

Dendrimers have found many applications in the fields of polymer science, biophysics, nanomedicine and the petroleum industry. Poly(amidoamine) (PAMAM) was studied as a model dendrimer and squalane as a model hydrocarbon. The interaction between PAMAM and squalane is pH dependent. Specifically, at low or neutral pH the squalane is found on the periphery of the PAMAM while at high pH the hydrocarbon is entrapped inside the PAMAM molecules.

Single-molecule fluorescence revealed that the interaction between PAMAM and squalane is reversible. At a pH value of 8, the time constants for the approaching, binding and dissociation of single PAMAM to ...


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 Jul 2008

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 (PAOX1) 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 a green ...


Spectroscopic And Molecular Dynamics Evidence For A Sequential Mechanisms For The A-To-B Transition In Dna, Ishita Mukerji Jul 2008

Spectroscopic And Molecular Dynamics Evidence For A Sequential Mechanisms For The A-To-B Transition In Dna, Ishita Mukerji

Division III Faculty Publications

The A-to-B form transition has been examined in three DNA duplexes, d(CGCGAATTCGCG)2, d(CGCGAATTGCGC), and d(CGCAAATTTCGC), using circular dichroism spectroscopy, ultraviolet resonance Raman (UVRR) spectroscopy, and molecular dynamics (MD) simulation. Circular dichroism spectra confirm that these molecules adopt the A form under conditions of reduced water activity. UVRR results, obtained under similar conditions, suggest that the transition involves a series of intermediate forms between A and B. Cooperative and distinct transitions were observed for the bases and the sugars. Independent MD simulations on d(CGCGAATTCGCG)2 show a spontaneous change from the A to B form in ...


In Vivo Murine Melanoma Tumor Responses To Nanosecond Pulsed Electric Field Treatment, Xinhua Chen Jul 2008

In Vivo Murine Melanoma Tumor Responses To Nanosecond Pulsed Electric Field Treatment, Xinhua Chen

Theses and Dissertations in Biomedical Sciences

High intensity nanosecond pulsed electric fields (nsPEF) were applied to melanoma tumors to observe functional and structural biological changes and to investigate the possible molecular mechanisms responsible. An animal model was set up by injecting B16F10 mouse melanoma cells into SKH-1 mice. A treatment (Tx) of 100 pulses: 300 nanosecond duration; 40 kV/cm field strength; at 0.5 Hz rate were delivered to melanoma tumors in 120 mice. The nsPEF Txcaused tumor self-destruction with sharply decreased cell volumes and shrunken nuclei. The apoptotic biochemical tests confirmed nsPEF Tx induced apoptosis in a time-dependent manner. Examination of gross vessel and ...


Pge2 Receptor Subsets Mediate Hcg Regulation Of The Plasminogen Activator/Plasmin System In Primate Periovulatory Follicles, Nune Markosyan Jul 2008

Pge2 Receptor Subsets Mediate Hcg Regulation Of The Plasminogen Activator/Plasmin System In Primate Periovulatory Follicles, Nune Markosyan

Theses and Dissertations in Biomedical Sciences

Elevated follicular prostaglandin E2 (PGE2) levels are necessary for follicle rupture during ovulation. As the degradation of the follicle wall requires proteolysis, it was hypothesized that regulation of proteolysis by gonadotropins is PGE2 mediated.

There are four PGE2 receptors: EP1, EP2, EP3 and EP4. To determine which PGE2 receptors are present in monkey periovulatory follicles, granulosa cells and whole ovaries were collected 0, 12, 24, and 36 hours after administration of an ovulatory dose of human chorionic gonadotropin (hCG), or 36 hours after hCG from monkeys that received the prostaglandin (PG) synthesis inhibitor celecoxib. All EP receptor mRNAs were present ...