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Preparation and Characterization of Model Conjugates for the Study of Proteins Modified by ADP-ribose

Description: Modification of proteins by ADP-ribose has been shown to be a versatile modification with respect to the amino acid side chain. The results described here will allow the study of the biological importance of ADP-ribose glycation and also allow differentiation on crude extracts between enzymatic modifications from protein ADP-ribose glycation that can occur due to the presence of NAD glycohydrolases.
Date: August 1992
Creator: Cervantes-Laurean, Daniel
open access

Identification and Characterization of an Arabidopsis thaliana Mutant with Tolerance to N-lauroylethanolamine

Description: N-Acylethanolamines (NAEs) are fatty acid derivatives in plants that negatively influence seedling growth. N-Lauroylethanolamine (NAE 12:0), one type of NAE, inhibits root length, increases radial swelling of root tips and reduces root hair numbers in a dose dependent manner in Arabidopis thaliana L. (ecotype Columbia). A forward genetics approach was employed by screening a population of T-DNA “activation-tagged” developed by the Salk Institute lines for NAE resistance to identify potential … more
Date: December 2015
Creator: Adhikari, Bikash
open access

Manipulations of Sucrose/Proton Symporters and Proton-pumping Pyrophosphatase Lead to Enhanced Phloem Transport But Have Contrasting Effects on Plant Biomass

Description: Delivery of photoassimilate, mainly sucrose (Suc) from photoautotrophic source leaves provides the substrate for the growth and maintenance of sink tissues such as roots, storage tissues, flowers and fruits, juvenile organs, and seeds. Phloem loading is the energized process of accumulating solute in the sieve element/companion cell complex of source leaf phloem to generate the hydrostatic pressure that drives long-distance transport. In many plants this is catalyzed by Suc/Proton (H+) symporte… more
Date: May 2015
Creator: Khadilkar, Aswad S
open access

Functional Significance of Sympathetic Fiber Ingrowth in the Habenula

Description: The physiological significance of noradrenergic sympathohabenular ingrowth following medial septal lesions was investigated. Following septal lesions, sympathetic fibers originating in the superior cervical ganglia are known to sprout into the medial habenular nuclei, and into the hippocampal formation. Previous work involving sympathohippocampal ingrowth showed that firing rates in septal animals with no ingrowth showed that firing rates in septal animals with no ingrowth were higher than rate… more
Date: August 1986
Creator: Howard, A. Jean (Ava Jean)
open access

Induced CSF-1 Production and its Effects on C-FMS Transfected Monoblastic U937 Cells

Description: This study examined how the monoblast-like human histiocytic lymphoma cell line U937 can be induced by phorbol 12-myristrate 13-acetate (PMA) to undergo differentiation. In order to study the mechanism of action of CSF-1, a CSF-1 receptor gene (c-fms) was transfected into U937 cells. Exogenous CSF-1 treatment induced an autocrine response in this CSF-1 was determined and all events were shown to be time dependent. CSF-1 stimulation also enhanced proto-oncogene c-jun and c-myc gene expression. … more
Date: August 1992
Creator: Liu, Mu-ya
open access

Studies on Poly(ADP-ribose) Metabolism and Chromatin Structure

Description: In these studies, a procedure which allowed the in vivo labeling and detection of poly(ADP-ribose) was combined with nuclear fractionation techniques to analyze the nuclear distribution of ADP-ribose polymers. The results from these studies suggest the occurrence of poly(ADP-ribose) metabolism in two compartments of chromatin; one that is nuclear matrix-associated and one that is not. The biological significance of this compartmentalization is conceptualization in a model. This model postulates… more
Date: August 1988
Creator: Cárdenas-Corona, María E. (María Elena)
open access

Occurrence and Structure of an Activating Enzyme for an S6 Kinase Determined by Monoclonal Antibody Analysis

Description: In this study, the production of monoclonal antibodies directed against the activating enzyme for an S6 kinase is examined and described. Evidence is presented for the association of an Mr. 55,000 abd Mr. 95,000 protein with the s6 kinase. These proteins are phosphorylated in the presence of Activating Enzyme. A sequence of regulatory events for insulin-stimulated phosphorylation of ribosomal protein S6 in cells is postulated as follows: insulin activates the receptor tyrosine kinase, which pho… more
Date: May 1987
Creator: Murdoch, Fern E. (Fern Elizabeth)
open access

Development of Enabling Technologies to Visualize the Plant Lipidome

Description: Improvements in mass spectrometry (MS)-based strategies for characterizing the plant lipidome through quantitative and qualitative approaches such as shotgun lipidomics have substantially enhanced our understanding of the structural diversity and functional complexity of plant lipids. However, most of these approaches require chemical extractions that result in the loss of the original spatial context and cellular compartmentation for these compounds. To address this current limitation, severa… more
Date: August 2013
Creator: Horn, Patrick J.
open access

Molecular and Functional Characterization of Medicago Truncatula Npf17 Gene

Description: Legumes are unique among plants for their ability to fix atmospheric nitrogen with the help of soil bacteria rhizobia. Medicago truncatula is used as a model legume to study different aspects of symbiotic nitrogen fixation. M. truncatula, in association with its symbiotic partner Sinorhizobium meliloti, fix atmospheric nitrogen into ammonia, which the plant uses for amino acid biosynthesis and the bacteria get reduced photosynthate in return. M. truncatula NPF1.7 previously called MtNIP/LATD is… more
Date: December 2013
Creator: Salehin, Mohammad
open access

Functional Characterization of Mtnip/latd’s Biochemical and Biological Function

Description: Symbiotic nitrogen fixation occurs in plants harboring nitrogen-fixing bacteria within the plant tissue. The most widely studied association is between the legumes and rhizobia. In this relationship the plant (legumes) provides the bacteria (rhizobia) with reduced carbon derived from photosynthesis in exchange for reduced atmospheric nitrogen. This allows the plant to survive in soil, which is low in available of nitrogen. Rhizobia infect and enter plant root and reside in organs known as nodul… more
Date: December 2013
Creator: Bagchi, Rammyani
open access

Modification of Cardiac Membrane Gsα by an Endogenous Arginine-Specific Mono-Adp-Ribosyltransferase

Description: The mechanism by which nicotinamide adenine dinucleotide (NAD) stimulates the activity of adenylate cyclase (AC) in canine plasma membrane has been studied. Using [3 2P]-NAD, the activation by NAD was correlated with the radiolabeling of the stimulatory guanosine triphosphate (GTP) binding protein Gsa. Further characterization demonstrated that the modification occurred only in the presence of G-protein activators and that arginine residue(s) were modified by ADP-ribose by the action of a mono-… more
Date: December 1993
Creator: Coyle, Donna L. (Donna Lynn)
open access

Physical, Chemical and Catalytic Properties of the Isozymes of Bovine Glucose Phosphate Isomerase

Description: Glucose phosphate isomerase (GPI) occurs in different bovine tissues as multiple, catalytically active isozymes which can be resolved by polyacrylamide gel electrophoresis and isoelectric focusing. GPI from bovine heart was purified to homogeneity and each of the isozymes was resolved. Four of the five isozymes were characterized with regard to their physical, chemical and catalytic properties in order to establish their possible physiological significance and to ascertain their molecular basis… more
Date: August 1987
Creator: Cini, John Kenneth
open access

Studies of Enzyme Mechanism Using Isotopic Probes

Description: The isotope partitioning studies of the Ascaris suum NAD-malic enzyme reaction were examined with five transitory complexes including E:NAD, E:NAD:Mg, E:malate, E:Mg:malate, and E:NAD:malate. Three productive complexes, E:NAD, E:NAD:Mg, and E:Mg:malate, were obtained, suggesting a steady-state random mechanism. Data for trapping with E:14C-NAD indicate a rapid equilibrium addition of Mg2+ prior to the addition of malate. Trapping with 14C-malate could only be obtained from the E:Mg2+:14C-malate… more
Date: August 1987
Creator: Chen, Cheau-Yun
open access

Purification, Characterization and Receptor Binding of Human Colony-Stimulating Factor-1

Description: Human colony-stimulating factor-1 (CSF-1) was purified from the serum-free conditioned medium of a human pancreatic carcinoma cell line. The four-step procedure included chromatography on DEAE Sepharose, Con A Sepharose and HPLC on phenyl column and reverse-phase C-3 column. The purity of human CSF-1 was demonstrated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS—PAGE) as a single diffuse band with a molecular weight (Mr) of 42,000-50,000 and was further confirmed by a single … more
Date: May 1987
Creator: Shieh, Jae-Hung
open access

Studies of the Mechanism of the Catalytic Subunit of cAMP Dependent Protein Kinase

Description: The kinetic mechanism of the cAMP-dependent protein kinase has been determined to be random in the direction of MgADP phosphorylation by using initial velocity studies in the absence and presence of the product, phospho-Serpeptide (Leu-Arg-Arg-Ala-Ser[P]-Leu-Gly) , and dead-end inhibitors. In contrast to the kinetic parameters obtained in the direction of Serpeptide phosphorylation, the only kinetic parameters affected by Mg^2+ are the dissociation constants for E:phospho-Serpeptide and E:MgADP… more
Date: August 1989
Creator: Yoon, Moon-Young
open access

Kinetic and Chemical Mechanism of Pyrophosphate-Dependent Phosphofructokinase

Description: Data obtained from isotope exchange at equilibrium, exchange of inorganic phosphate against forward reaction flux, and positional isotope exchange of 18O from the (βγ-bridge position of pyrophosphate to a (β-nonbridge position all indicate that the pyrophosphate-dependent phosphofructokinase from Propionibacterium freudenreichii has a rapid equilibrium random kinetic mechanism. All exchange reactions are strongly inhibited at high concentrations of the fructose 6-phosphate/Pi and MgPPi/Pi subst… more
Date: December 1988
Creator: Cho, Yong Kweon
open access

Studies on Hog Plasma Lecithin:cholesterol Acyltransferase: Isolation and Characterization of the Enzyme

Description: Lecithin:cholesterol acyltransferase (LCAT) was isolated from hog plasma and basic physicochemical properties and functionally important regions were investigated. Approximately one milligram of the enzyme was purified to apparent homogeneity with approximately a 20,000-fold increase in specific activity. In the plasma, hog LCAT was found to associate with high-density lipoproteins (HDL) probably through hydrophobic interactions with apolipoprotein A-I. HDL was the preferred lipoprotein substra… more
Date: May 1987
Creator: Park, Yong Bok
open access

Analysis of a Human Transfer RNA Gene Cluster and Characterization of the Transcription Unit and Two Processed Pseudogenes of Chimpanzee Triosephosphate Isomerase

Description: An 18.5-kb human DNA segment was selected from a human XCharon-4A library by hybridization to mammalian valine tRNAiAc and found to encompass a cluster of three tRNA genes. Two valine tRNA genes with anticodons of AAC and CAC, encoding the major and minor cytoplasmic valine tRNA isoacceptors, respectively, and a lysine tRNAcuu gene were identified by Southern blot hybridization and DNA sequence analysis of a 7.1-kb region of the human DNA insert. At least nine Alu family members were found inte… more
Date: August 1990
Creator: Craig, Leonard C. (Leonard Callaway)
open access

Studies of the Mechanism of Plasma Cholesterol Esterification in Aged Rats

Description: The study was performed to determine factors influencing the esteriflcation of plasma cholesterol in young and aged rats. The distribution of LCAT activity was determined following gel nitration chromatography and ultracentrifugation of whole plasma respectively. When rat plasma was fractionated on a Bio-Gel A-5 Mcolumn, LCAT activity was found to be associated with the HDL fraction. A similar result was observed upon 24 hr density gradient ultracentrifugation of the plasma. However, following … more
Date: December 1989
Creator: Lee, Sun Min
open access

Alternate Substrates and Isotope Effects as a Probe of the Malic Enzyme Reaction

Description: Dissociation constants for alternate dirmcleotide substrates and competitive inhibitors suggest that the dinucleotide binding site of the Ascaris suum NAD-malic enzyme is hydrophobic in the vicinity of the nicotinamide ring. Changes in the divalent metal ion activator from Mg^2+ to Mn^2+ or Cd^2+ results in a decrease in the dinucleotide affinity and an increase in the affinity for malate. Primary deuterium and 13-C isotope effects obtained with the different metal ions suggest either a change … more
Date: August 1988
Creator: Gavva, Sandhya Reddy
open access

Mechanism of the Adenosine 3',5'-Monophosphate Dependent Protein Kinase

Description: Isotope partitioning experiments were carried out with the adenosine 3',5'-monophosphate-dependent protein kinase catalytic subunit (cAPK) from bovine hearts to obtain information on the order of addition of reactants and the relative rates of reactant release from enzyme compared to the catalytic step(s). A value of 100% trapping for both ErMgATP-[γ-32P] and E:3H-Serpeptide at low Mgf indicates that MgATP and Serpeptide dissociate slowly from the enzyme compared to the catalytic step(s). The K… more
Date: May 1988
Creator: Kong, Cheng-Te
open access

Purification of HMG-CoA Reductase and Regulation by Protein-Lipid Interactions

Description: The enzyme 3-Hydroxy-3- Methylglutaryl Coenzyme A Reductase catalyzes the rate limiting step of hepatic cholesterol biosynthesis and is unique among the enzymes in the early part of the pathway in that it is membrane bound. This gives rise to potential regulation of the enzyme through interactions with the endoplasmic reticulum membrane. A purification procedure has been developed which consistently produces enzyme of high specific activity. In order to fully characterize the interactions betwe… more
Date: December 1982
Creator: Brent, Lynn G. (Lynn Gran)
open access

Analysis of Human Transfer RNA Gene Heteroclusters

Description: Two phage lambda clones encompassing human tRNA genes have been isolated from a human gene library harbored in bacteriophage lambda Charon-UA. One of the clones (designated as hLeuU) containing a 20-kb human DNA fragment was isolated and found to contain a cluster of four tRNA genes. An 8.2-kb Hindlll fragment encompassing the four tRNA genes was isolated from the 20-kb fragment and subcloned into pBR322 for restriction mapping and DNA sequence analysis. The four tRNA genes are arranged as two… more
Date: December 1986
Creator: Chang, Yung-Nien
open access

Metabolism of Diadenosine-5ʹ,5ʹʹʹ-P¹,P⁴-tetraphosphate (Ap₄A) in Cultured Mammalian Cells

Description: Methodology was developed which allowed the rapid and routine quantitation of subpicomole quantities of diadenosine-5ʹ,5ʹʹʹ-P¹,P⁴-tetraphosphate (Ap₄A) in cultured mammalian cells. This methodology includes the rapid extraction of cellular nucleotides in cold alkali, resolution of Ap₄A from the bulk of cellular materials on a highly specific boronate affinity resin, and quantitation of the dinucleotide in a coupled bioluminescence assay utilizing venom phosphodiesterase and firefly luciferase. … more
Date: December 1984
Creator: Baker, Jeffrey C. (Jeffrey Clayton)
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