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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)
Partner: UNT Libraries
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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
Partner: UNT Libraries
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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
Partner: UNT Libraries
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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
Partner: UNT Libraries
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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
Partner: UNT Libraries
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Structural Analyses of a Human Valine Transfer RNA Gene and of a Transfer RNA Pseudogene Cluster

Description: Two different cloned human DNA segments encompassing transfer RNA gene and pseudogene clusters have been isolated from a human gene library harbored in bacteriophage lambda Charon 4-A. One clone (designated as λhVal7) encompassing a 20.5-kilobase (Kb) human DNA insert was found to contain a valine transfer RNA_AAC gene and several Alu-like elements by Southern blot hybridization analysis and DNA sequencing with the dideoxyribonucleotide chain-termination method in the bacteriophage M13mp19 vect… more
Date: December 1987
Creator: Lee, Mike Ming-Jen
Partner: UNT Libraries
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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
Partner: UNT Libraries
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Purification and Characterization of a Differentiation Factor From Rat Lung Conditioned Medium

Description: A Differentiation Factor (DF) was purified from rat lung conditioned medium by a four-steps procedure. The DF has a molecular weight of 27000, and an isoelectric point of 4.70. Although DF is stable up to 60°C, it is sensitive to digestion by trypsin, chymotrypsin and subtilisin. DF forms granulocyte colonies in soft agar. Studies using anti-NRK CSF antibody demonstrated that DF is distinct from GM-CSF.
Date: May 1988
Creator: Ansari, Naser A. (Naser Awni)
Partner: UNT Libraries
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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
Partner: UNT Libraries
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In Vitro Modulation of Rat Liver Glyoxalase II Activity

Description: Glyoxylase II (Glo II, E.C. 3.1.2.6) catalyzes the hydrolysis of S-D-Lactoylglutathione (SLG) to D-Lactate and glutathione. This is the rate limiting step in the conversion of methylglyoxal to D-Lactate. The purpose of the present study was to determine whether or not a relationship exists between some naturally occuring metabolites and in vivo modulation of Glo II. We have observed a non-competitive inhibition (~ 45%) of Glo II in crude preparation of rat liver by GTP (0.3 mM). A factor (appar… more
Date: August 1988
Creator: Mbamalu, Godwin E.
Partner: UNT Libraries
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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)
Partner: UNT Libraries
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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
Partner: UNT Libraries
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Studies of the Interaction of LCAT with Lipoprotein Substrates in HDL Deficient Plasma Systems

Description: Enzymatic and lipid transfer reactions involved in reverse cholesterol transport were studied in HDL deficient plasma systems. Fasting plasma samples were obtained from control and cholesterol fed guinea pigs as well as from a fish eye disease patient and were used to localize the enzyme LCAT among plasma lipoproteins (VLDL, LDL, and HDL). In both guinea pig and fish eye disease patient plasma, the LCAT activity was found in association with the HDL type particles. Cholesterol feeding in guinea… more
Date: August 1989
Creator: Paranjape, Sulabha
Partner: UNT Libraries
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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
Partner: UNT Libraries
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Characterization of Human Glucose-6-Phosphate Isomerase of Different Sizes

Description: Glucose phosphate isomerase (GPI) was purified from human placenta utilizing cross-linked spherical particle phosphocellulose. In three steps, GPI could be purified approximately 5500 fold with greater than 50% recovery. The purified enzyme exhibited four bands upon non-denaturing PAGE and isoelectric focusing (IEF) when stained with GPI specific activity stain. The four isozymes were isolated by preparative IEF. The isoelectric points of the isozymes were determined. Sodium dodecyl sulfate (SD… more
Date: December 1989
Creator: Sun, An Qiang
Partner: UNT Libraries
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Physical Mapping of Human Transfer RNA Gene Clusters

Description: Two plaque-pure phage lambda clones designated as λhtX-l and λhtX-2 that hybridized to unfractionated bovine liver tRNA were isolated from a human X chromosome-specific library. The λDNAs were characterized by restriction mapping and Southern blot hybridization techniques. The human DNA segment in λhtX-l contains five or more presumptive tRNA genes and at least one Alu family member. The 19-kilobase human DNA insert in λhtX-2 contains two or more presumptive tRNA genes and at least three Alu fa… more
Date: December 1989
Creator: Wang, Luping
Partner: UNT Libraries
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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
Partner: UNT Libraries
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Sensitive Microtiter Assays for NAD, NADP and Protein Quantification in Human Lymphocytes

Description: Intracellular levels of NAD are of renewed interest in clinical and basic science research due to the new discovery of enzymes which utilize NAD as a substrate. Microtiter assays for the determination of NAD, NADP and protein were developed as modifications of previously published methods. The resulting assays are simple, cost effective and sensitive. An improved method of isolating lymphocytes was also developed. The resultant procedure requires one hour and removes greater than 99.9% of the p… more
Date: May 1990
Creator: Johnson, James, 1964-
Partner: UNT Libraries
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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)
Partner: UNT Libraries
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Studies on ADP-Ribose Polymer Metabolism in Cultured Mammalian Cells Following DNA Damage

Description: ADP-ribose polymer metabolism has been studied in human cells derived from a patient with Glutamyl Ribose Phosphate Storage Disease (GRPSD) and in mouse C3H1OT1/2 cells following oxidative stress induced by hydrogen peroxide (H202 ). It has been postulated that GRPSD resulted from an abnormality in ADP-ribose polymer metabolism. This study has shown that these cells exhibit reduced poly(ADP ribose) polymerase activity which is proposed to result from modification of the enzyme with ribose phosp… more
Date: May 1991
Creator: Maharaj, Geeta
Partner: UNT Libraries
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Chemical Mechanism of the Catalytic Subunit of Camp-Dependent Protein Kinase: Methods for Determining the Primary ¹⁸O Isotope Effects Using the Remote Label Technique

Description: A description of the nature of the transition state structure for phosphoryl transfer in the cAPK reaction requires a measurement of the primary 180 isotope effect at the serine hydroxyl acceptor. Since it is difficult to obtain primary 180 isotope effect directly, the 15N/1 4N ratio of the a-amine of the C-terminal glycine in the peptide Leu Arg-Lys-Ala-Ser-Leu-Gly (when serine is phosphorylated) was used to represent on the phosphorylation at serine. 15N Glycine, ' 4N-Glycine and 180 serine w… more
Date: December 1991
Creator: Chen, Gang, 1963-
Partner: UNT Libraries
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Evidence for the Interaction of GTP with Rat Liver Glyoxalase II

Description: Glyoxalase 11, the second enzyme of the glyoxalase system, hydrolyzes S-D-lactoylglutathione (SLG) to regenerate glutathione (GSH) and liberate free D-lactate. It was found that GTP binds with Gil from rat liver and inhibits Gil activity. Preincubation experiments showed that the binding is relatively tight, since more than 15 minutes are required to release GTP from the complex following dilution. Inhibition kinetics studies indicate that GTP is a "partially competitive inhibitor"; Thus, it wo… more
Date: December 1991
Creator: Yuan, Win-Jae
Partner: UNT Libraries
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Pre-Steady State Kinetics of the NAD-Malic Enzyme from Ascaris suum in the Direction of Oxidative Decarboxylation of L-Malate

Description: Stopped-flow experiments in which the NAD-malic enzyme was preincubated with different reactants at near saturating substrate concentrations suggest a slow isomerization of the E:NAD:Mg complex. The lag is eliminated by preincubation with Mg˙² and malate suggesting that the formation of E:Mg:Malate either bypasses or speeds up the slow isomerization step. Circular dichroic spectral studies of the secondary structural changes of the native enzyme in the presence and absence of substrates support… more
Date: December 1991
Creator: Rajapaksa, Ranjani, 1949-
Partner: UNT Libraries
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Studies on Poly (ADP-ribose) Synthesis in Lymphocytes of Systemic Lupus Erythematosus Patients

Description: A method for assaying poly (ADP-ribose) polymerase (PADPRP) activity in lymphocytes of systemic lupus erythematosus (SLE) patients has been developed. Using this method, PADPRP activity has been studied in lymphocytes from 15 patients and 13 controls. The mean activity in SLE lymphocytes was significantly lower than that in controls and 60% of the SLE patients demonstrated activities below the minimum of the control population. Possible mechanisms for this altered metabolism were investigated. … more
Date: December 1991
Creator: Chen, Hai-Ying
Partner: UNT Libraries
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