Biochemistry, Biophysics, and Structural Biology Commons^™

Direct Optical Scanning Gel Chromatography: Improvements To The Technique And Application To Several Diverse Biochemical Problems, John Grothusen

All Dissertations

No abstract provided.

A Study Of The Action Of Bovine Cathepsin D On Intramuscular Connective Tissue, Elimosaria Elikalia Maeda

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Cathepsin D has been postulated to play a role in the tenderization of meat with ageing. Meat toughness has been related to connective tissue content and degree of myofibrillar shortening. This investigation was undertaken to determine the activity of cathepsin D on bovine intramuscular connective tissue.

Highly purified cathepsin D preparations from bovine spleen and bovine sternomandibularis muscle were obtained by ammonium sulfate fractionation followed by affinity chromatography on pepstatin coupled to controlled pore size alkyl amine glass beads. Maximum proteolytic activity using hemoglobin as substrate was obtained at pH 3.5 and 3.8 for bovine spleen and muscle cathepsin D, …

Investigation Of The Substrate Binding Site Of Nad Glycohydrolase In Human Seminal Plasma, Sau C. Cheung

Chemistry & Biochemistry Theses & Dissertations

Nicotinamide adenine dinucleotide glycohydrolase (NADase) (E.C. 3.2.2.5) from human seminal plasma (HSP) was partially purified through a multistep purification procedure, including affinity chromatography on Amicon Red Matrex Gel A. The NADase activity was assayed by a fluorometric method with nicotinamide 1,N⁶ -ethenoadenine dinucleotide as the substrate. The enzymatic cleavage of etheno-NAD by NADase was accompanied by an eight-fold increase in fluorescence. This large increase in fluorescence provided a sensitive way to follow the rate of etheno-NAD hydrolysis.

NADase was found to be less specific for the purine moiety of NAD, since etheno-NAD was accepted as a substrate. The enzyme …

Conformational Analysis Of Nonsteroidal Estrogen Analogs, Robert N. Spady

Loma Linda University Electronic Theses, Dissertations & Projects

In target tissue, estrogens initiate a series of events, ending in protein synthesis, by binding to a cytoplasmic receptor. Once the estrogen binds, the receptor complex is activated--a process including dimerization of the estrogen complex with another subunit. In order to study the possible receptor-ligand interactions, conformational analysis was performed on several nonsteroidal estrogen analogs, using the CAMSEQ8 molecular mechanics program.

The probability that a molecule will exist in a given conformational energy well depends both on the energy and the shape of the well. A method has been previously worked out to calculate probabilities using both of these variables, …

Acetylation Of Synaptosomal Protein: Inhibition Of Veratridine / Soll Berl, Ramiro Nunez, Arlene D. Colon, And Donald D. Clarke Mount Sinai School Of Medicine, And Chemistry Department, Fordham University, New York, New York, U.S.A., Soll Berl, Ramiro Nunez, Arlene D. Colon, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Incubation of synaptosomes with [3H]acetate results in rapid labeling of protein. Labeling is decreased in the presence of veratridine, and the effect of veratridine is blocked by tetrodotoxin. Most of the radioactivity can be removed by base or acid hydrolysis, and is probably incorporated as acetate; it is this fraction that is affected by the veratridine. The data suggest that veratridine stimulates deacetylation of synaptosomal protein. This raises the question whether acetylation-deacetylation is involved in membrane function

The Metabolic Compartmentation Concept / S. Berl And D. D. Clarke Mt. Sinai School Of Medicine, Bronx, N.Y. 10458; Department Of Chemistry, Fordham University, New York, N.Y. 10029., Soll Berl, Donald Dudley Clarke Phd

Chemistry Faculty Publications

The concept of metabolic compartmentation describes the presence in a tissue of functionally different and chemically distinct pools of a given substrate. These separate pools equilibrate only very slowlyt if at a11, and exhibit different turnover and flux rates. Such heterogeneous functional pools of amino acids were coming under investigation in microorganisms (Britten et al. 1955; Cowie, Walton 1956; Cowie, McClure 1959), plants (Steward et al. 1956; Maclennan et al. 1963), and animal tissues (Korner, Tarver 1957; Green, Lowther 1959; Kipnis et al. 1961) at about the same time that we began our studies on glutamate-glutamine metabolism in brain. The …

Fluoroacetate As A Possible Marker For Glial Metabolism In Vivo / Donald D. Clarke, Ph.D. And Soll Berl, M. D. Dept. Of Chemistry, Fordham Univ. And Dept. Of Neurology, Mt. Sinai School Of Medicine Bronx, N.Y. 10458 And New York, N.Y. 10029, Donald Dudley Clarke Phd, Soll Berl, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Glucose is the major source of energy in all brain cells and its metabolism is closely coupled to functional activity. Advantage has been taken of this by Sokoloff (1977) and Sokoloff et al. (1977) to study energy metabolism in different brain areas and their response to a variety of drugs and stimuli. For this purpose these workers have developed the procedure for mrasuring autoradiographically the accumulation of [14C]-deoxyglucose into its phosphate derivative, a compound which does not differentiate glial from neuronal metabolism. They could be enhanced if one could differentiate glial from neuronal metabolic response to the various experimental conditions. …

Water Quality Trends In The Las Vegas Wash Wetlands, F. A. Morris, L. J. Paulson

Publications (WR)

The Las Vegas Wash is a wetlands ecosystem that acts to buffer the effects of wastewater discharges on the receiving waters of Lake Mead. The wash is the terminus for the 4,144 km2 Las Vegas Valley drainage basin, emptying into Las Vegas Bay of Lake Mead (Colorado River). It is in the northern Mojave desert, which receives an average of only 10 cm of rainfall annually. The Las Vegas Wash is technically an artificial wetland supported almost entirely by the perennial flows from sewage treatment plants. These flows contribute an average of 3-7 t of nutrients (nitrogen and phosphorus) and …

Biochemistry, W. E. Cornatzer

UND Departmental Histories

This departmental history was written on the occasion of the UND Centennial in 1983.

Maternal Ethanol Consumption: Glycoproteins And Gangliosides Of The Central Nervous System In The Rat Offspring, Antonio Blavatsky Caesar Noronha

Dissertations

No abstract provided.

Neutral Sugars In Selected Pit Viper, Elapid, Lizard And Scorpion Venoms, Ahmed El-Sherif, Paul D. Gwinup, Bob D. Johnson, Dewey H. Sifford

Journal of the Arkansas Academy of Science

No abstract provided.

The Relationship Between Whole Brain Catecholamine Depletion In Carassius Auratus And The Exposure To Inescapable Shock In A Learned Helplessness Paradigm, Roderick J. Misunis

Masters Theses

Learned helplessness is a psychological concept that describes the subsequent escape-avoidance behavior of experimental subjects who are exposed to uncontrollable stressors. Subjects after treatment are unable to respond in situations where escape is possible. Two major theorists, Seligman and Weiss, have proposed explanations concerning the phenomena. Seligman feels that the subject is unable to respond due to the fact that the exposure to an uncontrollable stressor has caused him to learn to be "helpless". Weiss feels that the inability to respond can best be described by alterations in the subject's brain neurochemistry, specifically the neurotransmitter norepinephrine. This paper is an …

Computer Pattern Recognition Of Action Potentials, Ronald S. Remmel

Journal of the Arkansas Academy of Science

A general method of pattern recognition was applied to the problem of recognizing extracellularly-recorded neuronal action potentials in the presence of noise and other pulses. A PDP1 1/23 performed the calculations. There were four stages: 1.) A bandpass filter attenuated noises; 2.) the data input program digitized the signal every 55 μsec. If the signal exceeded a threshold, 12 samples of the signal and the time were written onto the disk; 3.) the pulse discriminating program recognized an action potential by fitting the 12 points with this function: v(t) = (a + bt + ct² ) exp(-t/Ƭ). For each pulse …

Nickel: A Micronutrient Element For Hydrogen-Dependent Growth Of Rhizobium Japonicum And For Expression Of Urease Activity In Soybean Leaves, Robert V. Klucas, F. Joe Hanus, Sterling A. Russell, Harold J. Evans

Department of Biochemistry: Faculty Publications

Soybean plants and Rhizobium japonicum 122 DES, a hydrogen uptake-positive strain, were cultured in media purified to remove Ni. Supplemental Ni had no significant effect on the dry matter or total N content of plants. However, the addition of Ni to both nitrate-grown and symbiotically grown plants resulted in a 7- to 10-fold increase in urease activity (urea amidohydrolase, EC 3.5.1.5) in leaves and significantly increased the hydrogenase activity (EC 1.18.3.1) in isolated nodule bacteroids. When cultured under chemolithotrophic conditions, free-living R. japonicum required Ni for growth and for the expression of hydrogenase activity. Hydrogenase activity was minimal or not …

Reversible Dark-Induced Senescence Of Soybean Root Nodules, N. E. Pfeiffer, Nasir S.A. Malik, Fred W. Wagner

Department of Biochemistry: Faculty Publications

Nodule senescence was induced in intact soybean Glycine max. (L.) Merr., cv Woodworth plants by an 8-day dark treatment. Dark-induced senescence resulted in the complete loss of acetylene reduction activity, a 67% loss of total soluble protein, and an almost complete loss in total leghemoglobin of nodule extracts. Isoelectric focusing gels demonstrated a preferential loss of certain proteins, which was correlated with an increase in endoprotease specific activity toward azocasein. Nodules were completely green after the 8-day dark treatment. If plants were returned to a normal photoperiod after 8 days in the dark, nodules recovered from the dark treatment in …

Proteolytic Activity In Soybean Root Nodules: Activity In Host Cell Cytosol And Bacteroids Throughout Physiological Development And Senescence, Nancy E. Pfeiffer, Cecilia M. Torres, Fred W. Wagner

Department of Biochemistry: Faculty Publications

Root nodules were harvested from chamber-grown soybean (Glycine max L. Merril cv Woodworth) plants throughout development. Apparent nitrogenase activity (acetylene reduction) peaked before seeds began to develop, but a significant amount of activity remained as the seeds matured. Nodule senescence was defined as the period in which residual nitrogenase activity was lost. During this time, soluble protein and leghemoglobin levels in the host cell cytosol decreased, and proteolytic activity against azocasein increased. Degradative changes were not detected in bacteroids during nodule senescence. Total soluble bacteroid protein per gram of nodule remained constant, and an increase in proteolytic activity in …

Historical Patterns Of Phytoplankton Productivity In Lake Mead, Richard T. Prentki, Larry J. Paulson

Publications (WR)

Lake Mead was impounded in 1935 by the construction of Hoover Dam. The Colorado River was unregulated prior to then and therefore was subjected to extreme variations in flows and suspended sediment loads. Hoover Dam stabilized flows and reduced suspended sediment loads downstream, but Lake Mead still received silt-laden inflows from the upper Colorado River Basin. The Colorado River contributed 97% of the suspended sediment inputs to Lake Mead, and up to 140 x 1O⁶ metric tons (t) entered the reservoir in years of high runoff. Most of the sediments were deposited in the river channel and formed an …

Analysis Of The Surface Polysaccharides Of R. Trifolii Su843 And Its Sym Plasmid Alterated Mutants, Jauh-Lin Duh

Masters Theses

The Rhizobium trifolii polysaccharides from SU843 and mutants--HC843(pBR1AN), HC843, 8002--in which the sym plasmid is replaced, absent, or mutated by transposon Tn5 were isolated. The surface polysaccharides of genetically altered mutants were compared quantitatively and qualitatively with one another and with the parent strain. The interpretation of these results is discussed in relation to the function of Rhizobium surface polysaccharides in the symbiotic process. The symbiotic ability of SU843 and mutant strains is as follows: (a) SU843, nod⁺, fix⁺; (b) mutant 8002, roa⁺ (still attaches to root hair), hac^-, nod^-; (c) …

Effects Of Wastewater Discharges On Periphyton Growth In Lake Mead, Nevada-Arizona, Marsha Korb Morris

Publications (WR)

A study of the effects of secondary-treated wastewater on periphyton growth in Lake Mead, Nevada-Arizona was conducted from September 1979 to December 1980. Periphyton ash-free dry weight, chlorophyll-a, dominant species composition, and alkaline phosphatase activity were measured on fiberglass substrates. Substrates were incubated for two to four weeks in littoral and limnetic habitats. Physical and chemical variables and phytoplankton chlorophyll-a were measured concurrently.

Transparency increased with increasing distance from the discharge. Secchi depth ranged from 0.75 m at the discharge confluence (station 2) in August, to greater than 20 m at the most distant stations (stations 9 and 10) in …

Spermidine-Condensed Phi X174 Dna Cleavage By Micrococcal Nuclease: Torus Cleavage Model And Evidence For Unidirectional Circumferential Dna Wrapping., Kenneth A. Marx, Thomas C. Reynolds

Dartmouth Scholarship

Spermidine-condensed phi X174 replicative form (RF) II DNA was digested with micrococcal nuclease to yield seven identifiable DNA bands forming an arithmetic fragment-length series. The DNA monomer unit length was found to be 780 +/- 80 base pairs. This result is most consistent with a proposed model for micrococcal nuclease cleavage of a DNA torus organized by the unidirectional, circumferential wrapping of B-geometry DNA. By a topological consideration, the blunt-end-rod-fusion model for torus formation [Eickbush, T. H. & Moudrianakis, E. N. (1978) Cell 13, 295-306] is shown to be inconsistent with our empirical solution results. We propose a continuous, circumferential …

Fidelity Of Dna Polymerase In Vitro, Yannacos C. Philopoulos

Dissertations

The fidelity of DNA Polymerase I, isolated from B. licheniformis and B. stearothermophilus, was tested in the presence of carcinogens and anticancer compounds. All of these compounds inhibited (62.4-99.2%) the synthesis of DNA using Poly(dA-dT) as a template. The compounds were inhibitory at all concentration levels tested, 0.01, 1.0 and 100 mM. The fidelity of the enzyme appeared to be enhanced by the presence of these compounds, as indicated by misincorporation (50-70%), that is, the incorporation of incorrect nucleotide. Misincorporation in the presence of either carcinogens or anticancer compounds was less than that of control. The results from both organisms …

Development Of A Procedure For Analysis Of High Density Lipoprotein Subclasses, Chan Chin

Chemistry & Biochemistry Theses & Dissertations

Human serum high density lipoprotein subclasses, HDL₂ and HDL₃ , were isolated by preparative salt density gradient ultracentrifugation and further analyzed by electrophoresis on a 4 to 15% concentration gradient polyacrylamide gel.

The separation of the major classes of serum lipoproteins was achieved after a single ultracentrifugation for 272,000 g at 15°c in a swinging bucket rotor. High resolution concentration gradient gel electrophoresis was found to be particularly suitable for the separation of lipoproteins. The isolation of HDL by precipitation methods, heparin-manganese and dextran-magnesium have also been evaluated in this study, and were found not to completely separate …

Isomers Of 5.8s Ribosomal Rna, David Smith

Chemistry & Biochemistry Theses & Dissertations

When whole cell RNA was separated by high resolution PAG (i.e., polyacrylamide gel) electrophoresis, 5.8S rRNA migrated as three higher mobility intense bands, and three faint bands of lower mobility. The three intense bands represent conformational isomers which migrated as a single band under denaturing conditions on PAG. The three faint bands represent conformational isomers of a minor form of 5.8S rRNA which is elongated at the 5' terminus of the molecule. This minor form migrates as a single band of lower mobility than the major form when electrophoresed on a denaturing PAG. The minor form was determined to have …

Electroimmunoassay A New Competitive Protein Binding Assay Using Antibody Selective Electrodes, George Raymond Connell

Chemistry & Biochemistry Theses & Dissertations

The purpose of this study was to utilize concept of antibody selective electrodes to develop a new competitive protein binding assay. This is a new concept and potentially important analytical technique because it combines the advantage of RIA (sensitivity and selectivity) with advantages of electrical assays (speed and low cost).

Electroimmunoassay: A New Competitive Protein Binding Assay Using Antibody Selective Electrodes, George Raymond Connell

Chemistry & Biochemistry Theses & Dissertations

The purpose of this study was to utilize the concept of antibody selective electrodes to developed a new competitive protein binding assay. This is a new concept and a potentially important analytical technique because it combines the advantages of RIA (sensitivity and selectivity) with the advantages of electrical assays (speed and low cost). A conjugate of PGE₂ and dibenzo—18- crown-6 (a cation selective ionophore) was synthesized, and incorporated into a plastic membrane. The ionophore conjugate increased the selectivity of a polyvinyl chloride (PVC) membrane to monovalent cations. Transmembrane potential was altered by anti-PGE₂ antisera in a concentration dependent …

Development Of An On-Line Computer System For Enzyme Kinetics Studies, Jacqueline R. Thweatt

Chemistry & Biochemistry Theses & Dissertations

The PDP-11/10 minicomputer was interfaced to the Beckman 25 ultraviolet/visible spectrophotometer and this on-line computer system was applied to enzyme kinetics studies. This interface involved hardware equipment to locate, amplify, and transmit the analog signal to an analog-to-digital converter, and software to acquire and store the digital data. Data processing programs were used to calculate kinetic parameters and statistical information from the acquired data.

The Varian Cary 219 ultraviolet/visible spectrophotometer can be readily interfaced to the Apple II Plus microcomputer by hardware equipment which can be purchased from the manufacturer. Various software may also be purchased for use in enzyme …

The Isolation And Characterization Of A Novel Iron Protein From Desulfovibrio Gigas, Alan Kay Smith

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The isolation, purification, and partial characterization of a novel iron-containing protein from the sulfate-reducing anaerobic bacterium, Desulfovibrio gigas, is described. The highly insoluble protein was isolated from the cell debris following osmotic shock of the bacteria. The insoluble fraction consistently contained about 90% of the cell-associated iron. This protein was treated with acid, chelating agents, detergents and proteases in order to study their effect on the iron solubility. The results of elemental analysis of a crude protein preparation were 5.3% iron, 2.9% sulfur and 11 .9% nitrogen. An independent colorimetric iron analysis showed 6.4% iron. The iron could be …

Assay, Purification, And Characterization Of A Pantetheine Hydrolyzing Enzyme From Pig Kidney, Carl Thomas Wittwer

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A microsomal glycoprotein hydrolyzing pantetheine to pantothenate and cysteamine has been solubilized and purified to homogeneity as determined by sodium dodecylsulfate electrophoresis. Four rapid, independent assays of pantetheine hydrolysis are described and compared along with a method for localizing enzymatic activity on polyacrylamide gels. The enzyme is solubilized on exposure to butanol and purified by heat treatment, (NH4)₂SO₄ fractionation, hydrophobic chromatography, and hydroxyapatite chromatography. The glycoprotein, purified 5600-fold in 22% yield, has a specific activity of 14 μmoles pantothenate produced/min/mg of protein, 35 times that previously reported. The enzyme has a pH optimum of 9.0-9.5 and a …

Ortho Cyclization Reactions Of Catecholamines, Robert Lee Hudkins

Chemistry & Biochemistry Theses & Dissertations

A study was undertaken to examine the potential for ortho cyclization associated with the synthesis of 1,2,3,4,-tetrahydroisoquinolines using the Pictet-Spengler reaction. Specif1cally, the condensation of the biogenic amines, dopamine, norepinephrine and epinephrine were carefully examined in light of current literature and the facility and extent of ortho cyclization in each case. The para isomers of these three biogenic amine condensation products were isolated and characterized, while only the ortho isomer derived from dopamine was found stable and could be isolated, and structurally identified. These studies demonstrate that a second cyclization product does, in fact, form in each instance and that …

High-Temperature Battery Calorimeter, L. D. Hansen, R. H. Hart, D. M. Chen, H. F. Gibbard

Faculty Publications

A battery calorimeter was built for the measurement of thermal energy generation of high-temperature lithium–aluminum/iron sulfide battery cells, which are under development for electric vehicle propulsion and other energy storage applications. The calorimeter was designed with a temperature range of 400˚–500˚C, a detection limit of 1 mW, and an upper limit of heat flow of 50 W. The results of measurements on 200-Ah LiAl/FeS cells were in excellent agreement with the predictions of thermodynamic calculations based on precise measurements of the total cell polarization and the temperature coefficient of the emf. Details of the construction and operation principles of this …