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Articles 1 - 16 of 16

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

Protein-Protein And Protein-Dna Interactions At The Bacteriophage T4 Dna Replication Fork. Characterization Of A Fluorescently Labeled Dna Polymerase Sliding Clamp, Daniel J. Sexton, Theodore E. Carver, Anthony J. Berdis, Stephen J. Benkovic Nov 1996

Protein-Protein And Protein-Dna Interactions At The Bacteriophage T4 Dna Replication Fork. Characterization Of A Fluorescently Labeled Dna Polymerase Sliding Clamp, Daniel J. Sexton, Theodore E. Carver, Anthony J. Berdis, Stephen J. Benkovic

Chemistry Faculty Publications

The T4 DNA polymerase holoenzyme is composed of the polymerase enzyme complexed to the sliding clamp (the 45 protein), which is loaded onto DNA by an ATP-dependent clamp loader (the 44/62 complex). This paper describes a new method to directly investigate the mechanism of holoenzyme assembly using a fluorescently labeled cysteine mutant of the 45 protein. This protein possessed unaltered function yet produced substantial changes in probe fluorescence intensity upon interacting with other components of the holoenzyme. These fluorescence changes provide insight into the role of ATP hydrolysis in holoenzyme assembly. Using either ATP or the non-hydrolyzable ATP analog ...


The Carboxyl Terminus Of The Bacteriophage T4 Dna Polymerase Is Required For Holoenzyme Complex Formation, Anthony J. Berdis, Patrice Soumillion, Stephen J. Benkovic Nov 1996

The Carboxyl Terminus Of The Bacteriophage T4 Dna Polymerase Is Required For Holoenzyme Complex Formation, Anthony J. Berdis, Patrice Soumillion, Stephen J. Benkovic

Chemistry Faculty Publications

To further elucidate the mechanism and dynamics of bacteriophage T4 holoenzyme formation, a mutant polymerase in which the last six carboxyl-terminal amino acids are deleted, was constructed, overexpressed, and purified to homogeneity. The mutant polymerase, designated ΔC6 exo−, is identical to wild-type exo− polymerase with respect to kcat, kpol, and dissociation constants for nucleotide and DNA substrate. However, unlike wild-type exo− polymerase, the ΔC6 exo− polymerase is unable to interact with the 45 protein to form the stable holoenzyme. A synthetic polypeptide corresponding to the carboxyl terminus of the wild-type exo− polymerase was tested as an in vitro inhibitor of ...


Open-Path Fourier Transform Infrared Studies Of Large-Scale Laboratory Biomass Fires, Robert J. Yokelson, David W. T. Griffith, Darold E. Ward Sep 1996

Open-Path Fourier Transform Infrared Studies Of Large-Scale Laboratory Biomass Fires, Robert J. Yokelson, David W. T. Griffith, Darold E. Ward

Chemistry and Biochemistry Faculty Publications

A series of nine large-scale, open fires was conducted in the Intermountain Fire Sciences Laboratory (IFSL) controlled-environment combustion facility. The fuels were pure pine needles or sagebrush or mixed fuels simulating forest-floor, ground fires; crown fires; broadcast burns; and slash pile burns. Mid-infrared spectra of the smoke were recorded throughout each fire by open path Fourier transform infrared (FTIR) spectroscopy at 0.12 cm−1 resolution over a 3 m cross-stack pathlength and analyzed to provide pseudocontinuous, simultaneous concentrations of up to 16 compounds. Simultaneous measurements were made of fuel mass loss, stack gas temperature, and total mass flow up ...


High-Power Broadly Tunable Picosecond Ir Laser System For Use In Nonlinear Spectroscopic Applications, D. E. Gragson, B. M. Mccarty, G. L. Richmond, D. S. Alavi Sep 1996

High-Power Broadly Tunable Picosecond Ir Laser System For Use In Nonlinear Spectroscopic Applications, D. E. Gragson, B. M. Mccarty, G. L. Richmond, D. S. Alavi

Chemistry and Biochemistry

We developed a high-power tunable picosecond IR laser system suitable for nonlinear spectroscopic investigations. We employ a Ti:sapphire regenerative amplifier that produces 1.9-ps, 17-cm−1, 800-nm pulses at 1 kHz as a pump source. White-light generation in ethylene glycol and optical parametric amplification in potassium titanyl phosphate are used to produce the IR pulses. The tuning range extends from 2.4 to 3.8 μm in the idler and 1.0 to 1.2 μm in the signal. A total efficiency (signal plus idler) as high as 20% was achieved. The spatial, spectral, and temporal characteristics of the ...


Critical Pressures In Multicomponent Lipid Monolayers, John P. Hagen, Harden M. Mcconnell Apr 1996

Critical Pressures In Multicomponent Lipid Monolayers, John P. Hagen, Harden M. Mcconnell

Chemistry and Biochemistry

Epifluorescence microscopy has been used previously to study coexisting liquid phases in lipid monolayers of dihydrocholesterol and dimyristoylphosphatidylcholine at the air/water interface. This binary mixture has a critical point at room temperature (22°C), a monolayer pressure of approx. 10 mN/m, and a composition in the vicinity of 20-30 mol% dihydrocholesterol. It is reported here that this critical pressure can be lowered, raised, or maintained constant by systematically replacing molecules of this phosphatidylcholine with molecules of a phosphatidylethanolamine, or an unsaturated phosphatidylcholine, or mixtures of the two, while maintaining the dihydrocholesterol concentration at 20 mol%. Thus, even complex ...


The Lipooligosaccharides Of Haemophilus Ducreyi Are Highly Sialylated, William Melaugh, A A. Campagnari, B W. Gibson Jan 1996

The Lipooligosaccharides Of Haemophilus Ducreyi Are Highly Sialylated, William Melaugh, A A. Campagnari, B W. Gibson

Chemistry Faculty Publications

The major lipooligosaccharides of the sexually transmitted pathogen Haemophilus ducreyi 35000 have been previously found to terminate in N-acetyllactosamine and sialyl-N-acetyllactosamine, Neu5Ac alpha 2-->3Gal beta 1-->4GlcNAc (W. Melaugh, N. J. Phillips, A. A. Campagnari, M. V. Tullius, and B. W. Gibson, Biochemistry 33: 13070-13078, 1994). In this study, mass spectrometry and composition analyses have shown that the lipooligosaccharides from three other H. ducreyi strains also contain N-acetyllactosamine and are highly sialylated (approximately 30 to 50%), although one African strain was found to contain neither of these structural features.


Degradation Studies Of The Non-Lethal Bird Repellent, Methyl Anthranilate, Eugeny Aronov, Larry Clark Jan 1996

Degradation Studies Of The Non-Lethal Bird Repellent, Methyl Anthranilate, Eugeny Aronov, Larry Clark

Larry Clark

Methyl anthranilate (MA), a food grade flavor and fragrance additive, has been reported to be an effective non-lethal bird repellent in a variety of situations. Despite the experimental success of MA, field studies have yielded widely differing levels of efficacy. Diminished efficacy in some field trials prob­ ably results from the failure of specific formulations to retain or protect the active ingredient under natural conditions. Therefore, a clearer understanding of the physical and chemical factors affecting the stability of MA is needed. We undertook a series of laboratory studies on hydrolysis, photolysis and microbial degradation of MA, the results of ...


An Improved Synthesis Of 2-(Hydroxymethyl)Indene, Hasan Palandoken, William T. Mcmillen, Michael H. Nantz Jan 1996

An Improved Synthesis Of 2-(Hydroxymethyl)Indene, Hasan Palandoken, William T. Mcmillen, Michael H. Nantz

Chemistry and Biochemistry

No abstract provided.


Characterization Of Biomacromolecules By Nuclear Magnetic Resonance Spectroscopy , Wayne Robert Baker Jan 1996

Characterization Of Biomacromolecules By Nuclear Magnetic Resonance Spectroscopy , Wayne Robert Baker

Retrospective Theses and Dissertations

Two proteins were examined by Nuclear Magnetic Resonance (NMR) spectroscopy. The Emb-1 protein of Daucus carota was found to have few spectral characteristics indicative of a stable tertiary structure. In particular, line-widths were narrow, chemical shift dispersion was poor and nuclear Overhauser effect spectra were sparse. The NMR spectra of Ribonuclease A was examined at a range of pH values and the chemical shifts of a selected number of resonances was plotted as a function of the pH. From these titrations curves pKa values were determined and assigned to specific ionizing groups within the protein based, in part, on physical ...


Applications Of Partial Filling Micellar Electrokinetic Chromatography: From Electrospray Mass Spectrometry To Fluorescence Polarization Immunodetection , Wendy Marie Nelson Jan 1996

Applications Of Partial Filling Micellar Electrokinetic Chromatography: From Electrospray Mass Spectrometry To Fluorescence Polarization Immunodetection , Wendy Marie Nelson

Retrospective Theses and Dissertations

Mechanistic studies of partial filling micellar electrokinetic chromatography PF-MEKC) and a theoretical model for the prediction of separation behavior in PF-MEKC was developed in this work. Along with the theoretical development of PF-MEKC, two different detection modes were applied to PF-MEKC, in contrast to the traditional UV detection used in micellar electrokinetic chromatography (MEKC). The first detection mode coupled PF-MEKC with eletrospray ionization mass spectrometry. The second detection mode developed in this work is PF-MEKC coupled with fluorescence polarization immunodetection;First, in the mechanistic studies of PF-MEKC a theoretical model was proposed for predicting the separation behavior of triazine herbicides ...


Single Cell Analysis By Capillary Electrophoresis With Laser-Induced Native Fluorescence Detection , Sheri Joanne Lillard Jan 1996

Single Cell Analysis By Capillary Electrophoresis With Laser-Induced Native Fluorescence Detection , Sheri Joanne Lillard

Retrospective Theses and Dissertations

Individual mammalian cells were analyzed by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). This technique was chosen due to its high separation efficiencies, small sample volumes and sensitive detection. Native fluorescence was used, in which the analyte was not tagged with a fluorophore. 275-nm excitation gave attomole (amol = 10-18 mol) detection limits for the intracellular species of interest. Two projects are described in which hemoglobin (Hb) variants were determined in single red blood cells. In the third experiment, individual mast cells were degranulated on-column, and exocytosis and serotonin release were monitored temporally;First, single red blood cells, in which Hb ...


A New High Pressure Sapphire Nuclear Magnetic Resonance Cell, Shi Bai, Craig M. Taylor, Charles L. Mayne, Ronald J. Pugmire, David M. Grant Jan 1996

A New High Pressure Sapphire Nuclear Magnetic Resonance Cell, Shi Bai, Craig M. Taylor, Charles L. Mayne, Ronald J. Pugmire, David M. Grant

Faculty Publications

A new version of a single-crystal sapphire high pressure nuclear magnetic resonance (NMR) cell is described that is capable of controlling the sample pressure independent of the temperature. A movable piston inside the cell adjusts and controls the sample pressure from ambient conditions to 200 atm within plus or minus 0.3 atm. The linewidth at half-height for a 13C spectrum of carbon dioxide at 15degrees C and 57.8 atm is found to be 0.5 Hz. The carbon dioxide gas/liquid phase transition is clearly observed by measuring 13C chemical shifts as the sample pressure approaches equilibrium. The ...


Mechanistic Studies On Malate Dehydrogenase From Escherichia Coli, S. Kirk Wright Jan 1996

Mechanistic Studies On Malate Dehydrogenase From Escherichia Coli, S. Kirk Wright

Masters Theses

Kinetic studies and chemical modification studies were performed on malate dehydrogenase from Escherichia coli (eMDH). Chemical modification studies using diethylpyrocarbonate and iodoacetic acid, along with log V/K profiles indicate that one ionizing group with a pKa between 7.9 and 8.5 acts as a general acid/base in the catalytic mechanism. Log V profiles suggest that malate binds to the correctly protonated form of the enzyme. These results imply that a histidine residue is required for catalysis.

A primary deuterium kinetic isotope effect of 1.43 (±0.14) was observed on V/K using malate-2-d, while no ...


Space-Based Genetic Cryoconservation Of Endangered Species, Michael Noah Mautner Jan 1996

Space-Based Genetic Cryoconservation Of Endangered Species, Michael Noah Mautner

Chemistry Publications

Genetic materials of endangered species must be maintained, for cryoconservation, permanently near liquid nitrogen temperatures below 77 K. Due to the instability of human institutions, permanent safety is best provided at storage sites that maintain passively the needed low temperatures, and provide barriers to access. The required conditions are available in permanently shaded polar lunar craters with equilibrium temperatures of 8 to 40 K, on the moons of Saturn, and unshielded storage satellites. A genetic depository can be incorporated readily into planned lunar programmes.


In-Situ Imaging Of Ionic Crystal Dissolution Using An Integrated Electrochemical/Afm Probe, Julie V. Macpherson, Patrick R. Unwin, Andrew C. Hillier, Allen J. Bard Dec 1995

In-Situ Imaging Of Ionic Crystal Dissolution Using An Integrated Electrochemical/Afm Probe, Julie V. Macpherson, Patrick R. Unwin, Andrew C. Hillier, Allen J. Bard

Andrew C. Hillier

The kinetics and mechanism controlling dissolution from the (100) cleavage face of potassium bromide single crystals in acetonitrile solutions have been identified using a novel integrated electrochemical/AFM probe and a scanning electrochemical microscope (SECM). With both techniques, dissolution is induced by perturbing the dynamic dissolution/growth equilibrium at the crystal/solution interface through the electrochemical oxidation of bromide ions. SECM measurements demonstrate that the dissolution reaction is diffusion-limited under the experimental conditions, suggesting that the surface reaction is characterized by a rate constant in excess of 5 cm s-1 (assuming a first-order dissolution process). The topography of the dissolving ...


Measurement Of Double-Layer Forces At The Electrode/Electrolyte Interface Using The Atomic Force Microscope: Potential And Anion Dependent Interactions, Andrew C. Hillier, Sunghyun Kim, Allen J. Bard Dec 1995

Measurement Of Double-Layer Forces At The Electrode/Electrolyte Interface Using The Atomic Force Microscope: Potential And Anion Dependent Interactions, Andrew C. Hillier, Sunghyun Kim, Allen J. Bard

Andrew C. Hillier

The forces between a silica probe and silica and gold substrates were measured with an atomic force microscope in the presence of a series of alkali-halide electrolyte solutions. The interaction between two silica surfaces was repulsive and could be accurately predicted by Derjaguin-Landau-Verwey-Overbeek theory. The silica surface was negatively charged at a pH of 5.5 and the effective surface potential increased in magnitude with decreasing electrolyte concentration. In contrast, the force between the silica probe and a gold substrate was attractive at open circuit. This interaction was a strong function of the potential applied to the gold and the ...