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Articles 1 - 29 of 29

Full-Text Articles in Cell Biology

Serum Deprivation Of Mesenchymal Stem Cells Improves Exosome Activity And Alters Lipid And Protein Composition, Reka A. Haraszti, Rachael Miller, Michelle L. Dubuke, Andrew H. Coles, Marie C. Didiot, Dimas Echeverria, Matteo Stoppato, Yves Y. Sere, John D. Leszyk, Julia F. Alterman, Bruno M. D. C. Godinho, Matthew R. Hassler, Rachel Wollacott, Yan Wang, Scott A. Shaffer, Neil Aronin, Anastasia Khvorova May 2019

Serum Deprivation Of Mesenchymal Stem Cells Improves Exosome Activity And Alters Lipid And Protein Composition, Reka A. Haraszti, Rachael Miller, Michelle L. Dubuke, Andrew H. Coles, Marie C. Didiot, Dimas Echeverria, Matteo Stoppato, Yves Y. Sere, John D. Leszyk, Julia F. Alterman, Bruno M. D. C. Godinho, Matthew R. Hassler, Rachel Wollacott, Yan Wang, Scott A. Shaffer, Neil Aronin, Anastasia Khvorova

Open Access Articles

Exosomes can serve as delivery vehicles for advanced therapeutics. The components necessary and sufficient to support exosomal delivery have not been established. Here we connect biochemical composition and activity of exosomes to optimize exosome-mediated delivery of small interfering RNAs (siRNAs). This information is used to create effective artificial exosomes. We show that serum-deprived mesenchymal stem cells produce exosomes up to 22-fold more effective at delivering siRNAs to neurons than exosomes derived from control cells. Proteinase treatment of exosomes stops siRNA transfer, indicating that surface proteins on exosomes are involved in trafficking. Proteomic and lipidomic analyses show that exosomes derived in ...


Serinc5: Its Sensitivity To Nef And Restriction Of Hiv-1, Weiwei Dai Aug 2018

Serinc5: Its Sensitivity To Nef And Restriction Of Hiv-1, Weiwei Dai

GSBS Dissertations and Theses

The accessory protein Nef of human immunodeficiency virus type 1 (HIV-1) has long been known to enhance the infectivity of HIV-1 progeny virions. The multipass transmembrane proteins serine incorporator 3 (SERINC3) and SERINC5 were recently identified as novel antiviral proteins that restrict HIV-1 infectivity. Nef enhances HIV-1 infectivity by removing SERINCs from the plasma membrane, which prevents their incorporation into progeny HIV-1 virions. To exploit this potent intrinsic antiretroviral factor for potential therapy development, it is critical to explore the determinants in SERINC5 that govern its downregulation by Nef and its restriction on HIV-1 infectivity. Here I report that the ...


Regulation Of The Pi3-Kinase/Pten Signaling Pathway By Tgf-Β In Prostate Cancer Cells, Mawiyah Kimbrough-Allah May 2018

Regulation Of The Pi3-Kinase/Pten Signaling Pathway By Tgf-Β In Prostate Cancer Cells, Mawiyah Kimbrough-Allah

Electronic Theses & Dissertations Collection for Atlanta University & Clark Atlanta University

Transforming growth factor -β (TGF-β) plays an important role in the progression of prostate cancer. It acts as a tumor suppressor in normal epithelial cells but as a tumor promoter in advanced prostate cancer cells. The PI3-kinase pathway has been shown to play integral roles in many cellular processes including cell proliferation, survival, and cell migration in many cell types. PI3-kinase pathway mediates TGF-β effects on prostate cancer cell migration and invasion. Phosphatase and tensin homolog (PTEN), a tumor suppressor gene, inhibits PI3-kinase pathway and is frequently mutated in prostate cancers. In this present study, we investigated possible roles of ...


Heavily And Fully Modified Rnas Guide Efficient Spycas9-Mediated Genome Editing, Aamir Mir, Julia F. Alterman, Matthew R. Hassler, Alexandre J. Debacker, Edward Hudgens, Dimas Echeverria, Michael H. Brodsky, Anastasia Khvorova, Jonathan K. Watts, Erik J. Sontheimer Mar 2018

Heavily And Fully Modified Rnas Guide Efficient Spycas9-Mediated Genome Editing, Aamir Mir, Julia F. Alterman, Matthew R. Hassler, Alexandre J. Debacker, Edward Hudgens, Dimas Echeverria, Michael H. Brodsky, Anastasia Khvorova, Jonathan K. Watts, Erik J. Sontheimer

University of Massachusetts Medical School Faculty Publications

RNA-based drugs depend on chemical modifications to increase potency and nuclease stability, and to decrease immunogenicity in vivo. Chemical modification will likely improve the guide RNAs involved in CRISPR-Cas9-based therapeutics as well. Cas9 orthologs are RNA-guided microbial effectors that cleave DNA. No studies have yet explored chemical modification at all positions of the crRNA guide and tracrRNA cofactor. Here, we have identified several heavily-modified versions of crRNA and tracrRNA that are more potent than their unmodified counterparts. In addition, we describe fully chemically modified crRNAs and tracrRNAs (containing no 2'-OH groups) that are functional in human cells. These designs ...


Near-Infrared Photoactivatable Control Of Ca Signaling And Optogenetic Immunomodulation, Lian He, Yuanwei Zhang, Guolin Ma, Peng Tan, Zhan Jun Li, Shengbing Zhang, Xiang Wu, Ji Jing, Shaohai Fang, Lijuan Zhou, Youjun Wang, Yun Huang, Patrick Hogan, Gang Han, Yubin Zhou Dec 2015

Near-Infrared Photoactivatable Control Of Ca Signaling And Optogenetic Immunomodulation, Lian He, Yuanwei Zhang, Guolin Ma, Peng Tan, Zhan Jun Li, Shengbing Zhang, Xiang Wu, Ji Jing, Shaohai Fang, Lijuan Zhou, Youjun Wang, Yun Huang, Patrick Hogan, Gang Han, Yubin Zhou

Open Access Articles

The application of current channelrhodopsin-based optogenetic tools is limited by the lack of strict ion selectivity and the inability to extend the spectra sensitivity into the near-infrared (NIR) tissue transmissible range. Here we present an NIR-stimulable optogenetic platform (termed "Opto-CRAC") that selectively and remotely controls Ca2+ oscillations and Ca2+-responsive gene expression to regulate the function of non-excitable cells, including T lymphocytes, macrophages and dendritic cells. When coupled to upconversion nanoparticles, the optogenetic operation window is shifted from the visible range to NIR wavelengths to enable wireless photoactivation of Ca2+-dependent signaling and optogenetic modulation of immunoinflammatory responses. In a ...


Development And Application Of Mass Spectrometry-Based Proteomics To Generate And Navigate The Proteomes Of The Genus Populus, Paul Edward Abraham May 2013

Development And Application Of Mass Spectrometry-Based Proteomics To Generate And Navigate The Proteomes Of The Genus Populus, Paul Edward Abraham

Doctoral Dissertations

Historically, there has been tremendous synergy between biology and analytical technology, such that one drives the development of the other. Over the past two decades, their interrelatedness has catalyzed entirely new experimental approaches and unlocked new types of biological questions, as exemplified by the advancements of the field of mass spectrometry (MS)-based proteomics. MS-based proteomics, which provides a more complete measurement of all the proteins in a cell, has revolutionized a variety of scientific fields, ranging from characterizing proteins expressed by a microorganism to tracking cancer-related biomarkers. Though MS technology has advanced significantly, the analysis of complicated proteomes, such ...


Novel Role Of Antioxidant-1 (Atox1) As A Copper-Dependent Transcription Factor Involved In Cell Proliferation, S. Itoh, H. W. Kim, O. Nakagawa, K. Ozumi, Susan M. Lessner, H. Aoki, K. Akram, R. D. Mckinney, M. Ushio-Fukai, T. Fukai Feb 2008

Novel Role Of Antioxidant-1 (Atox1) As A Copper-Dependent Transcription Factor Involved In Cell Proliferation, S. Itoh, H. W. Kim, O. Nakagawa, K. Ozumi, Susan M. Lessner, H. Aoki, K. Akram, R. D. Mckinney, M. Ushio-Fukai, T. Fukai

Faculty Publications

Copper plays a fundamental role in regulating cell growth. Many types of human cancer tissues have higher copper levels than normal tissues. Copper can also induce gene expression. However, transcription factors that mediate copper-induced cell proliferation have not been identified in mammals. Here we show that antioxidant-1 (Atox1), previously appreciated as a copper chaperone, represents a novel copper-dependent transcription factor that mediates copper-induced cell proliferation. Stimulation of mouse embryonic fibroblasts (MEFs) with copper markedly increased cell proliferation, cyclin D1 expression, and entry into S phase, which were completely abolished in Atox1-/- MEFs. Promoter analysis and EMSA revealed that copper stimulates ...


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 ...


Hras Intracellular Trafficking And Signal Transduction, Jodi Ho-Jung Mckay Jan 2007

Hras Intracellular Trafficking And Signal Transduction, Jodi Ho-Jung Mckay

Retrospective Theses and Dissertations

Cancer, which is characterized by uncontrolled cell division and proliferation, is the second leading cause of death in the United States. Of these deaths, approximately 30% are the result of mutations in Ras protein. When activated, Ras propagates multiple signal transduction cascades and induces cell proliferation, differentiation, and apoptosis. Within the Ras subfamily, there are several isoforms of Ras which differ in their posttranslational modifications. The modifications include proteolytic cleavage of the CAAX box, farnesylation, and palmitoylation which act as plasma membrane anchors where most signal transduction takes place. This dissertation demonstrates that Ras endocytosis takes place through a novel ...


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 ...


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 ...


From Endomembrane To The Plasma Membrane: The Traffic, Signal Transduction And Micro-Localization Of H-Ras , Hui Zheng Jan 2006

From Endomembrane To The Plasma Membrane: The Traffic, Signal Transduction And Micro-Localization Of H-Ras , Hui Zheng

Retrospective Theses and Dissertations

Ras proteins are small G proteins that play key roles in many aspect of cell signal transduction. H-Ras is one of three isoforms of mammalian Ras protein that is best studied. The work presented in this dissertation describes how the carboxyl terminal lipid modifications affect the transportation, signal transduction and micro-localizations of H-Ras protein. The already known traffic pathway for H-Ras to the plasma membrane is Golgi mediated classical vesicular Transportation; However, we found the H-Ras transportation has many different features other than the classical pathway, and thereby discovered a new pathway for H-Ras targeting to the plasma membrane. This ...


Analysis Of A Nuclear Complex Of Proteins That Form A Spindle Matrix During Mitosis In Drosophila , Uttama Rath Jan 2005

Analysis Of A Nuclear Complex Of Proteins That Form A Spindle Matrix During Mitosis In Drosophila , Uttama Rath

Retrospective Theses and Dissertations

A spindle matrix is a macromolecular structure hypothesized to provide a stationary support or strut for the anchorage of motor proteins during force generation, microtubule sliding, and chromosome segregation in mitosis. Previous work in Drosophila has shown Skeletor, as the first putative molecular candidate of spindle matrix structure since it possesses many features consistent with the proposed spindle matrix structure.;We have identified two more putative spindle matrix candidate proteins, Chromator and Megator, using yeast two-hybrid interaction assay, cross-immunoprecipitation and immunocytochemistry. Chromator is a novel and essential chromodomain containing protein that interacts directly with Skeletor and extensively colocalizes with it ...


Investigation Of The Function Of Branching Enzymes I, Iia, And Iib In The Determination Of Amylopectin Structure And Regulation Of Starch Biosynthesis , Rebekah Suzanne Marsh Jan 2005

Investigation Of The Function Of Branching Enzymes I, Iia, And Iib In The Determination Of Amylopectin Structure And Regulation Of Starch Biosynthesis , Rebekah Suzanne Marsh

Retrospective Theses and Dissertations

Starch is one of the most abundant carbohydrates on earth serving vital roles as the primary energy reserve of plants, a significant source of calories in the human diet, and a renewable resource for other industrial applications. The multiplicity of starch biosynthetic enzymes is evolutionarily conserved in plants suggesting their functions are essential in starch biosynthesis. The overall goal of this research is to determine the specific molecular functions of the multiple starch branching enzyme (BE) isoforms in maize.;This dissertation first addresses what, if any, are the differences in enzymatic properties of BEIIa and BEIIb. Two polyclonal antisera were ...


Structural And Molecular Analysis Of Leech Neuronal Proteins, Calsensin And Filamin , Deepa V. Venkitaramani Jan 2004

Structural And Molecular Analysis Of Leech Neuronal Proteins, Calsensin And Filamin , Deepa V. Venkitaramani

Retrospective Theses and Dissertations

This thesis describes the structural and molecular analysis of two leech neuronal proteins, Calsensin and Filamin. Calsensin is an EF-hand calcium-binding protein expressed by a subset of peripheral sensory neurons which fasciculate into a single tract in the leech central nervous system. Calsensin is a 9 kD protein with two EF-hand calcium-binding motifs. Calsensin can multimerize via disulfide bridge formation involving the two cysteine residues in vitro. Furthermore, Calsensin shows differential gel migration depending on the presence or absence of calcium suggesting a conformational change upon calcium binding. Using multidimensional NMR spectroscopy we have determined the solution structure and backbone ...


The Essential Jil-1 Kinase Regulates Histone H3 Phosphorylation And Maintains Chromatin Structure In Drosophila , Weiguo Zhang Jan 2004

The Essential Jil-1 Kinase Regulates Histone H3 Phosphorylation And Maintains Chromatin Structure In Drosophila , Weiguo Zhang

Retrospective Theses and Dissertations

JIL-1 encodes a chromatin-associated tandem serine/threonine kinase in Drosophila melanogaster. JIL-1 predominantly associates with interbands on polytene chromosomes. It appears at higher levels on the X chromosome than on autosomes in males, and distributes about equally on all chromosomes in females. In addition, in males JIL-1 overlaps with the MSL (for male specific lethal) complex that specifically associates with the male X chromosome on which expression of most genes are upregulated.;We isolated JIL-1 mutants by inducing imprecise excision of an EP element inserted in JIL-1. We found that JIL-1 is required for viability of both females and males ...


Mechanisms Of Negative Regulation Of Protein Prenylation: Investigation Of Murine Guanylate-Binding Protein 1 , John Thomas Stickney Jan 2000

Mechanisms Of Negative Regulation Of Protein Prenylation: Investigation Of Murine Guanylate-Binding Protein 1 , John Thomas Stickney

Retrospective Theses and Dissertations

Covalent attachment of isoprenoid lipids to proteins is a post-translational modification that occurs on approximately 2% of all cellular proteins. Proteins that contain a CaaX motif as their final four amino acids are presumed to be targets for the addition of C15 or C20 isoprenoids by farnesyl transferase (FTase) or geranylgeranyl transferase I (GGTase I) respectively. This modification is generally believed to occur stoichiometrically, and irreversibly, in vivo. Thus, if prenylation is to be regulated, this control must occur prior to lipid addition. At the moment, there is little evidence that the enzymatic activity of prenyl transferases can be modulated ...


The Contributions Of The Hras C-Terminus And Its Lipid Modifications To Hras Signal Transduction Events And Membrane Binding , Michelle Ann Booden Jan 2000

The Contributions Of The Hras C-Terminus And Its Lipid Modifications To Hras Signal Transduction Events And Membrane Binding , Michelle Ann Booden

Retrospective Theses and Dissertations

In the GTP bound state, Ras proteins activate multiple downstream effector proteins, the combination of which regulate cell growth, differentiation, and changes in cell shape. However, Ras proteins must undergo C-terminal lipidation before they can support biological activity. Specifically, the C-terminus of the HRas protein must be modified by a farnesyl at Cys186 and two palmitates at Cys181 and Cys184. The current paradigm suggests that palmitate is not attached without prior farnesyl addition and that these post-translational lipid modifications are only necessary for membrane association. However, emerging evidence suggests that the C-terminus and its lipid modifications may also be responsible ...


Effects Of Altering Titin Expression On Myofibril Assembly In A Skeletal Muscle Cell Line , Qunfeng Dong Jan 2000

Effects Of Altering Titin Expression On Myofibril Assembly In A Skeletal Muscle Cell Line , Qunfeng Dong

Retrospective Theses and Dissertations

Titin is a giant muscle protein that spans the distance from the Z line to the M line in sarcomeres of striated muscle myofibrils. One of the functions proposed for titin is that it could act as a molecular scaffold to direct sarcomere assembly. To investigate this role of titin, titin expression was transiently altered in the cultured mouse skeletal muscle cell line C2C12 by either a dominant-negative approach or an antisense mRNA approach. The time course of myofibril assembly was monitored by immunofluorescence labeling with antibodies to actin, alpha-actinin, titin, and myosin from 1 to 5 d after myoblast ...


Fatty Acid Modification Of Cells In Culture By Overexpression Of Exogenous Desaturases , Travis Jay Knight Jan 1999

Fatty Acid Modification Of Cells In Culture By Overexpression Of Exogenous Desaturases , Travis Jay Knight

Retrospective Theses and Dissertations

COS-1 cells were transfected transiently with expression vectors encoding stearoyl-acyl-carrier-protein desaturase (SACPD) from castor bean, stearoyl-CoA desaturase (SCD) from mice, and a control vector containing SACPD cDNA in reverse orientation. The hypothesis was that both exogenous desaturases would be active in COS-1 cells, thereby increasing the proportion of monounsaturated fatty acids produced by the cells. Messenger RNA and anti-myc immunoreactive proteins were detected at 12 and 24 hours post-transfection respectively and increased through 48 hours post-transfection. When acetate was used as the fatty acid precursor in cultures for a 24-hour period starting 24 hours post-transfection, the ratio of monounsaturated to ...


Endogenous Alkylglycerol Functions As A Mediator Of Protein Kinase C Activity And Cell Proliferation, Fritz G. Buchanan May 1997

Endogenous Alkylglycerol Functions As A Mediator Of Protein Kinase C Activity And Cell Proliferation, Fritz G. Buchanan

Electronic Theses and Dissertations

To explore the possibility that 1-O-alkyl-sn-glycerol (alkylglycerol) may serve a regulatory role in the control of cell proliferation or PKC activity, we examined the ability of alkylglycerol to influence PKC activity and subcellular distribution as well as the ability of alkylglycerol to effect cell proliferation. MDCK cells grown to confluence show a loss of PKC activity associated with the membrane, as reported in fibroblasts. Preconfluent cultures of MDCK cells have a high level of PKC activity associated with the membrane. However, treatment of preconfluent cultures with alkylglycerol causes a reduction of PKC activity. A similar inhibition was observed with alkylglycerol ...


Identification Of Mouse Sip24/24p3 As A New Acute Phase Protein And Study Of Its Expression , Quansheng Liu Jan 1996

Identification Of Mouse Sip24/24p3 As A New Acute Phase Protein And Study Of Its Expression , Quansheng Liu

Retrospective Theses and Dissertations

Mouse SIP24 was originally discovered as a 24 k Da inducible protein secreted from quiescent Balb/c 3T3 fibroblast cells. Based on the analysis of peptide sequencing results and Western blot studies, SIP24 was identified as identical to the mouse protein encoded by 24P3 and is thus referred to as SIP24/24P3 in this thesis. SIP24/24P3 is a member of the lipocalin protein family, and its human homolog NGAL (neutrophil gelatinase associated lipocalin) has been reported to bind the chemotactic factor, fMLP (N-formylmethionyl-leucyl-phenylalanine);Studies were carried out to test the hypothesis that SIP24/24P3 is an acute phase protein ...


The Interaction Between Actin Filaments And The Cytoskeleton-Membrane Attachment Site Protein Talin , Jinwen Zhang Jan 1995

The Interaction Between Actin Filaments And The Cytoskeleton-Membrane Attachment Site Protein Talin , Jinwen Zhang

Retrospective Theses and Dissertations

Talin, a protein localized primarily in cell-matrix type adherens junctions, is involved in linking the cytoskeleton to the cell membrane. The major objectives of my study were to (1) determine the structural basis for the effect of pH and ionic strength on the direct talin-actin interaction, (2) examine the effects of other factors, such as specific phospholipids, on the talin-actin interaction, and (3) compare abilities of human platelet and avian smooth muscle talins to interact with actin. Negative staining results showed that at pH 6.4 and low ionic strength, talin extensively crosslinked actin filaments into both tight networks and ...


The Role(S) Of Arginine-Specific Mono(Adp-Ribosyl)Transferase In Skeletal Muscle Cells , Hui-Yu Huang Jan 1995

The Role(S) Of Arginine-Specific Mono(Adp-Ribosyl)Transferase In Skeletal Muscle Cells , Hui-Yu Huang

Retrospective Theses and Dissertations

The overall goal of this research project was to examine the biological function of arginine-specific mono(ADP-ribosyl) transferase in skeletal muscle cells. To better understand the function of this enzyme in skeletal muscle, the experimental approach to accomplish this goal consisted of two specific objectives. The first objective was to identify the protein substrates for this enzyme in myogenic cell cultures and cellular fractions. Embryonic chick primary muscle cell cultures were used as a working system for this purpose. In membrane fractions of 96-h myotube cultures incubated with [[superscript]32 P]NAD under conditions selected for mono(ADP-ribosyl)ation, proteins ...


5-Fluoroorotic Acid-Selected Cell Lines And Regulation Of Ump Synthase Gene Expression In Tobacco Cells , Djoko Santoso Jan 1995

5-Fluoroorotic Acid-Selected Cell Lines And Regulation Of Ump Synthase Gene Expression In Tobacco Cells , Djoko Santoso

Retrospective Theses and Dissertations

Plant uridine-5'-monophosphate (UMP) synthase has been studied at both the biochemical and molecular levels. Using a purification procedure of salt precipitation coupled with a series of chromatography techniques, UMP synthase was purified to more than 2,000-fold from crude extracts of Cucurbita pepo L. As with UMP synthases from other higher eukaryotes, the UMP synthase of squash has both orotate phosphoribosyl transferase and orotidine-5'-monophosphate (OMP) decarboxylase activities. The molecular mass of the native UMP synthase was about 110 kDa and SDS-PAGE showed a subunit molecular mass of 50 kDa. The other biochemical properties of this UMP synthase are ...


Examination Of The Effects Of Selected Factors On The Talin-Actin Interaction , Jean Marie Schmidt Jan 1994

Examination Of The Effects Of Selected Factors On The Talin-Actin Interaction , Jean Marie Schmidt

Retrospective Theses and Dissertations

Talin is an adhesion plaque protein believed to be involved in linking actin filaments to cell membranes. In these studies I examined the effects of selected factors on the ability of talin and its 190 kDa proteolytic fragment to interact with actin. A modified method for purifying talin from avian smooth muscle was devised. Cosedimentation assays showed that increasing amounts of talin bound to actin filaments as pH was decreased from 7.4 to 6.4. Talin's ability to increase viscosity of F-actin solutions also was increased with decreasing pH within this range. Electron microscope observations supported a pH-dependent ...


Mouse Mast Cell Proteases: Induction, Molecular Cloning, And Characterization, Wei Chu May 1991

Mouse Mast Cell Proteases: Induction, Molecular Cloning, And Characterization, Wei Chu

Electronic Theses and Dissertations

Tryptase, a mast cell-specific serine protease with trypsin-like specificity, has been identified in a mouse mast cell line (ABFTL-6) based on it's enzymatic activity, inhibition properties, and cross-reactivity to a human mast cell tryptase antibody. The effects of fibroblast-conditioned medium and sodium butyrate on ABFTL-6 mast cell differentiation and tryptase expression have been examined. ABFTL-6 mouse mast cells undergo phenotypic changes upon culturing in media supplemented with fibroblast-conditioned media at 50% or 1 mM sodium butyrate. The induced cells increased in size, had larger and more metachromatic cytoplasmic granules, and increased their total cellular protein about four-fold. Tryptase activity ...


Characterization Of The Mitogen Regulated Protein Receptor , Joseph T. Nelson Jan 1991

Characterization Of The Mitogen Regulated Protein Receptor , Joseph T. Nelson

Retrospective Theses and Dissertations

Mitogen regulated protein (MRP) is a glycoprotein synthesized in murine placenta and secreted into the maternal bloodstream during gestation. The temporal profile of MRP concentration in the plasma during gestation, its synthesis in an organ known to secrete other lactogenic hormones, and its homology to prolactin suggest an hormonal role;A requisite characteristic of a polypeptide hormone is to have a receptor on the plasma membrane of another tissue separate from its site of synthesis. The receptor then mediates the functional effects by binding the hormone. I discovered and characterized a receptor for MRP from murine uterine membrane preparations. The ...