Biochemistry, Biophysics, and Structural Biology Commons^™

Domains And Anomalous Adsorption Isotherms Of Dipalmitoylphosphatidylcholine Membranes And Lipophilic Ions: Pentachlorophenolate, Tetraphenylborate, And Dipicrylamine, Pavel Smejtek, S. Wang

Physics Faculty Publications and Presentations

Dipalmitoylphosphatidylcholine (DPPC) vesicles acquire negative surface charge on adsorption of negatively charged pentachlorophenolate (PCP-), and lipophilic ions tetraphenylborate (TPhB-), and dipicrylamine (DPA-). We have obtained (a) zeta-potential isotherms from the measurements of electrophoretic mobility of DPPC vesicles as a function of concentration of the adsorbing ions at different temperatures (25–42 degrees C), and (b) studied the effect of PCP- on gel-to-fluid phase transition by measuring the temperature dependence of zeta-potential at different PCP- concentrations. The zeta-potential isotherms of PCP- at 25, 32, and 34 degrees C correspond to adsorption to membrane in its gel phase. At 42 degrees C the …

Nitrogen Dioxide Reaction With Proteins: Evidence For Peptide Bond Cleavage At Lysine Residues, Darryl B. Hood

Electronic Theses and Dissertations

Nitrogen dioxide (NO$\sb2$), an air pollutant produced by burning fossil fuels and a component of cigarette smoke, is thought to contribute to the pathogenesis of pulmonary diseases, such as emphysema. To gain information on the mechanism by which NO$\sb2$ damages the lung, in vitro exposures of $\alpha\sb1$-proteinase inhibitor ($\alpha\sb1$-PI), elastin, bovine serum albumin (BSA), human serum albumin (HSA) and synthetic poly-L-lysine were performed. A genetic deficiency of $\alpha\sb1$-PI predisposes humans to emphysema and NO$\sb2$ has been hypothesized to damage $\alpha\sb1$-PI, which would leave proteases such as human neutrophil elastase, (HNE) free to attack lung structural proteins. The ability of $\alpha\sb1$-PI …

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 increased …

Metabolism Of Arachidonate-Containing Phospholipid Molecular Species In The Murine Macrophage-Like Cell Line, P388d1, Crystal R. Waites

Electronic Theses and Dissertations

Glycerophospholipids of mammalian cells exist as chemically diverse structures with various fatty acids at the sn-1 and sn-2 positions. Arachidonic acid, a polyunsaturated fatty acid, which may be converted to biologically active eicosanoids such as prostaglandins, thromboxanes, and leukotrienes, is found predominantly in the sn-2 position of glycerophospholipids. The purpose of this study was to examine, at the level of the individual molecular species, the incorporation of arachidonate into phospholipids and its release from phospholipids during stimulation. In this way, the specificity of the enzymes controlling arachidonate metabolism could be examined in order to clarify the processes that control the …

A Biochemical Study Of Gossypol And Lactate Dehydrogenase X Binary Interactions, Patricia Brown Ravenell

Theses and Dissertations in Biomedical Sciences

The goal of this project was to study the mechanism of action of gossypol through its (a) binary interaction with native and trypsin digested lactate dehydrogenase-X (LD-X), a sperm-specific isozyme, and (b) its binding in vitro in primary cultures of spermatogenic cells. Mouse LD-X was cleaved with trypsin before and after treatment with gossypol. This was followed by high performance chromatography (HPLC) separation of the LD-X tryptic peptide fragments to observed alterations in separation patterns as indicators of intramolecular disturbance in the enzyme molecule. Definite alterations in peptide fragment peaks were observed in the presence of gossypol and suggest conformational …

The Role Of Queuine Deficient Trna In Translational Control, Kathryn Lenore Hart

Theses and Dissertations in Biomedical Sciences

The purpose of this research was to study the effect of the level of tRNA modification with respect to queuine on translational efficiency. Transfer RNA from neoplastic cells is generally deficient with regard to certain hypermodified nucleosides found in normal cells. The hypothesis that was examined is whether or not the altered efficiency of translation can be correlated to the presence of queuine hypomodified tRNA. The loss of queuine in the anticodon may change the interaction of the tRNA's anticodon with its codon. Therefore, the translational complex may stall looking for a rare tRNA, or the stability of the complex …

The Study Of Natural Product(S) From Opsanus Tau (Linnaeus), The Oyster Toadfish, Miki Taira

Chemistry & Biochemistry Theses & Dissertations

Opsanus tau (Linnaeus), the Oyster toadfish, commonly inhabits the Chesapeake Bay. The Oyster toadfish has been categorized as a venomous and/or ichthyotoxic fish; however, chemical studies to substantiate these claims have not been forthcoming.

The oyster toadfish has dorsal and opercular spines which are believed to secrete a venom(s). Based on our findings, the oyster toadfish does not secrete venom(s) from these spines.

The oyster toadfish secretes large amounts of a mucus when it is irritated. 3-Octanone was believed to be the major ichthyotoxic component of the mucus from the oyster toadfish. Based on our investigation, 3-octanone is present only …

Molecular Mechanism Of The Stimulation Of Alkaline Phosphatase Activity In Human Bone Cells By 1,25(Oh)2 D3, Eru Kyeyune-Nyombi

Loma Linda University Electronic Theses, Dissertations & Projects

To facilitate this study an in vitro human model system was established that exhibited many aspects of normal osteoblasts. The human osteosarcoma cell line (TE85 cells) expressed a skeletal alkaline phosphatase activity (an accepted bone cell differentiation marker) which was stimulated by 1,25(OH)₂D₃ (a potent differentiating agent), under serum-free conditions in a dose-dependent, time-dependent, and cell density-dependent manner. Cytochemical analysis of the stimulation of ALP activity by 1,25(OH)₂D₃ showed that 1,25(OH)₂D₃ increased the number of TE85 cells that expressed detectable ALP activity, suggesting that 1,25(OH)₂D₃ promoted the process …

Cells That Express All Five Proteins Of Vesicular Stomatitis Virus From Cloned Cdnas Support Replication, Assembly, And Budding Of Defective Interfering Particles, Asit K. Pattnaik, Gail W. Wertz

School of Veterinary and Biomedical Sciences: Faculty Publications

An alternative approach to structurefunction analysis of vesicular stomatitis virus (VSV) gene products and their interactions with one another during each phase of the viral life cycle is described. We showed previously by using the vaccinia viruslT7 RNA polymerase expression system that when cells expressing the nucleocapsid protein (N), the phosphoprotein (NS), and the large polymerase protein (L) of VSV were superinfected with defective interfering (DI) particles, rapid and efflicient replication and amplification of DI particle RNA occurred. Here, we demonstrate that all five VSV proteins can be expressed simultaneously when cells are cotransfected with plasmids containing the matrix protein …

Interleukin-1 Or Phorbol Induction Of The Stromelysin Promoter Requires An Element That Cooperates With Ap-1., Karen Sirum-Connolly, Constance E. Brinckerhoff

Dartmouth Scholarship

Interieukin-1, a mediator of inflammation, or tumor promoting phorbol esters induce transcription of stromelysin, a metalloproteinase that degrades extracellular matrix molecules and that is overexpressed in diseases such as rheumatoid arthritis. Sequences required for induction of transcription of the human stromelysin promoter are contained on a 46 base pair fragment. This fragment contains a sequence with a high degree of similarity to the binding site for the transcription factor activator protein-1 (AP-1) and indeed, the AP-1 sequence of this fragment is necessary but not sufficient for the maximal response to phorbol 12-myristate 13-acetate (phorbol) or interleukin-1. Maximal induction requires functional …

Use Of Affinity Chromatography In Developing Acridinium Ester-Labeled Antibodies For An Immunometric Assay Of Parathyrin, David S. Hage, Bob Taylor, Pat Schryver, Pai C. Kao

David Hage Publications

In developing an immunometric assay of intact parathyrin (parathyroid hormone, PTH), we found that affinity chromatography is a useful tool in purifying and optimizing the labeling conditions for acridinium ester-labeled antibodies. ... In summary, affinity chromatography was found to be useful in the purification of acridinium ester-labeled antibodies, particularly for removing denatured antibodies from the prepared label and for monitoring the amount of active labeled antibodies produced.

Improved Syntheses Of Some Monochloro- And Monobromo-8-Quinolinols / Herman Gershon And Donald D. Clarke Department Of Chemistry, Fordham University, Bronx, Ny 10458, Usa, Herman Gershon, Donald Dudley Clarke Phd

Chemistry Faculty Publications

Procedures were developed for the preparation of the 2-, 3-, 4-, and 6-monosubstituted chloro and bromo 8-quinolinols which afforded greater yields and/or reduced the number of steps in the preparation. 100 MHz 1H-NMR spectra for the 12 possible monochloro and mono bromo analogues are given

Fluoroacetate And Fluorocitrate: Mechanism Of Action / Donald D. Clarke, Donald Dudley Clarke Phd

Chemistry Faculty Publications

The concept of lethal synthesis as suggested by Peters is reviewed in the light of the more recent work in this area. It is suggested that fluorocitrate is a "suicide" substrate for aconitase rather than a competitive inhibitor as originally suggested. The use of these substances to study glialneuronal relationships is considered

Evidence Of Steric Factors In The Fungitoxic Mechanism Of 8-Quinolinol And Its 5- And 7-Halogenated Analogues / Herman Gershon, Donald D. Clarke, And Muriel Gershon, Herman Gershon, Donald Dudley Clarke Phd, Muriel Gershon

Chemistry Faculty Publications

Antifungal studies were made of mixtures of minimal inhibitory concentrations (MICs) of 8-quinolinol and its 5- and 7-halo analogues against six fungi: Aspergillus niger, A. oryzae, Trichoderma viride, Myrothecium verrucaria, Mucor cirinelloides, and Trichophyton mentagrophytes. Mixtures of 8-quinolinol with 5- or 7-fluoro-8-quinolinol and of 5- and 7-fluoro-8-quinolinol showed additive activity, and their respective toxicities were reversed by L-cysteine. These results suggested a common mechanism of activity for the three toxicants. Potentiation of the fungitoxicity of mixtures of 8-quinolinol and its 5- and 7-chloro, bromo, and iodo analogues, as well as mixtures of 5- and 7-chloro, 5- and 7-bromo, and 5- …

Ganglioside Metabolism In Embryonic Chick Neurons, Michael Victor Hogan

Dissertations

No abstract provided.

Induction Of Gene Expression In Response To Stress In Pseudomonas Aeruginosa Pao, Audrey L. Warner-Bartnicki

Dissertations

No abstract provided.

Cell Surface-Binding Sites For Progesterone Mediate Calcium Uptake In Human Sperm, Peter F. Blackmore, Joseph Neulan, Frank Lattanzio, Stephen J. Beebe

Bioelectrics Publications

Recent studies (e.g. Blackmore, P. F., Beebe, S. J., Danforth, D. R., and Alexander, N.) (1990) J. Biol. Chem. 265, 1376-1380) have shown that in human sperm, progesterone produces a rapid increase in intracellular free calcium ([Ca2+]i) and an induction of the acrosome reaction (e.g. Osman, R. A., Andria, M. L., Jones, A. D., and Meizel, S. (1989) Biochem. Biophys. Res. Commun. 160, 828-833). In this study, the location of progesterone receptors on the cell surface of human sperm was identified using progesterone immobilized on bovine serum albumin (BSA) (progesterone 3-(O-carboxymethyl)oxime:BSA) as well as progesterone and its 3-O-carboxymethyloxime derivative. Using …

Purification And Characterization Of Soybean Root Nodule Ferric Leghemoglobin Reductase, Lin Ji, Stephen Wood, Manuel Becana, Robert V. Klucas

Department of Biochemistry: Faculty Publications

A ferric leghemoglobin reductase from the cytosol of soybean (Glyclne max) root nodules was purified to homogeneity and partlafly characterized. The enzyme is a flavoprotein with flavin adenine dinuclotide as the prosthetic group and consists of two identical subunits, each having a molecular mass of 54 kilodaltons. The pure enzyme shows a high activity for ferric leghemoglobin reduction with NADH as the reductant in the absence of any exogenous mediators. The enzyme also exhibits NADH-dependent 2,6-dichloroindophenol reductase activity. A sequence of the first 50 N-terminal amino acids of the purified protein was obtained. Comparisons with known protein sequences …

Detection Of Mercuric Ions In Water By Elisa With A Mercury- Specific Antibody, Dwane E. Wylie, Larry D. Carlson, Randy Carlson, Fred W. Wagner, Sheldon M. Schuster

Department of Biochemistry: Faculty Publications

An immunoassay that detects mercuric ions in water at concentrations of 0.5 ppb and above is described. The assay utilizes a monoclonal antibody that binds specifically to mercuric ions immobilized in wells of microtiter plates. Within the range of 0.5-10 ppb mercury, the absorbance in the enzyme-linked immunosorbent assay (ELISA) is linear to the log of the mercuric ion concentration. The quantitation of mercury by ELISA correlates closely with results from cold-vapor atomic absorption. Other divalent metal cations do not interfere with the assay, although there is interference in the presence of 1 mM chloride ions. The optimum pH for …

Immunochemical Characterization Of The Antigenic And Allergenic Components Of The Plant Pollens Of Paspalum Notatum. Callistemon Citrinis And Melaleuca Leucadendron, Saeed S. Hosseiny

Retrospective Theses and Dissertations

Antigenic as well as allergenic cross-reactivity was observed among the pollen extracts of Paspalum notatum, bahia (BA); Callistemon citrinis, bottlebrush (BO); and Melaleuca leucadendron, melaleuca (ME). lmmunoblot analysis of SOS-PAGE separated proteins revealed multiple antigenic components in the extracts of all three allergens which cross-reacted with rabbit antisera made to each pollen extract (anti-BA, anti-BO and anti-ME). A unique antigenic component of 15 kD molecular size reacting only to rabbit anti-BA antisera was detected in the bahia extract. Allergenic cross-reactivity among these pollens was strongly suggested by results obtained with skin testing of allergic individuals. A clinical study of 55 …

Photosynthetic Electron Transport In Genetically Altered Photosystem Ii Reaction Centers Of Chloroplasts, Robin A. Roffey, John H. Golebeck, C. Russ Hille, Richard T. Sayre

Department of Biochemistry: Faculty Publications

Using a cotransformation system to identify chloroplast transformants in Chlamydomonas reinhardtii, we converted histidine-195 of the photosystem H reaction center D1 protein to a tyrosine residue. The mutants were characterized by a reduced quantum efficiency for photosynthetic oxygen evolution, which varied in a pH-dependent manner, a reduced capacity to oxidize artificial donors to photosystem II, and P680⁺) reduction kinetics (microsecond) that were essentially similar to wild type. In addition, a dark-stable radical was detected by ESR in mutant photosystem II particles but not in wild-type particles. This radical was similar in g value and lineshape to chlorophyll …

The Effects Of Chronic Ethanol Exposure Of The Dopaminergic System Of The Rat Brain, Susan Marie Pellegrino

Dissertations

No abstract provided.

Monoclonal Antibodies To The Fusion Protein Of Bovine Respiratory Syncytial Virus, Kent M. Mulkey, Gary A. Anderson

School of Veterinary and Biomedical Sciences: Faculty Publications

Five monoclonal antibodies specific for bovine respiratory syncytial virus were characterized by Western immunoblotting, radioimmunoprecipitation, and epitope mapping assays. The monoclonal antibodies were found to be specific for the fusion protein, and there were at least two antigen binding sites, one of which was neutralizing.

Protein Turnover As A Component In The Light/Dark Regulation Of PhosphoEnolPyruvate Carboxylase Protein-Serine Kinase Activity In C4 Plants, Jin-An Jiao, Cristina Echevarría, Jean Vidal, Raymond Chollet

Department of Biochemistry: Faculty Publications

Maize leaf phosphoenolpyruvate carboxylase [PEPC; orthophosphate:oxaloacetate carboxy-lyase (phosphorylating), EC 4.1.1.311 protein-serine kinase (PEPC-PK) phosphorylates serine-15 of its target enzyme, thus leading to an increase in catalytic activity and a concomitant decrease in malate sensitivity of this cytoplasmic C₄ photosynthesis enzyme in the light. We have recently demonstrated that the PEPC-PK activity in maize leaves is slowly, but strikingly, increased in the light and decreased in darkness. In this report, we provide evidence that cycloheximide, an inhibitor of cytoplasmic protein synthesis, when fed to detached leaves of C₄ monocots (maize, sorghum) and dicots (Portulaca oleracea) in the …

Nicotinate, Nicotinamide, And The Reactivity Of Leghemoglobin In Soybean Root Nodules, Robert V. Klucas, Cyril A. Appleby

Department of Biochemistry: Faculty Publications

Nicotinate has been postulated to interfere with the binding of O₂ to ferrous leghemoglobin in soybean (Glycine max) root nodules. For such a function, the levels of nicotinate in nodules must be sufficiently high to bind a significant amount of leghemoglobin. We have measured levels of nicotinate, nicotinamide, and leghemoglobin in soybean nodules from plants 34 to 73 days after planting in a glasshouse. On a per gram nodule fresh weight basis, levels between 10.4 and 21 nanomoles for nicotinate, 19.2 and 37.8 nanomoles for nicotinamide, and 170 to 280 nanomoles for leghemoglobin were measured. Even if …

Posttranslational Regulation Of Phosphoenolpyruvate Carboxylase In C4 And Crassulacean Acid Metabolism Plants, Jin-An Jiao, Raymond Chollet

Department of Biochemistry: Faculty Publications

Control of C₄ photosynthesis and Crassulacean acid metabolism (CAM) is, in part, mediated by the diel regulation of phosphoenolpyruvate carboxylase (PEPC) activity. The nature of this regulation of PEPC in the leaf cell cytoplasm of C₄ and CAM plants is both metabolite-related and posttranslational. Specifically, the regulatory properties of the enzyme vary in accord with the physiological activity of C₄ photosynthesis and CAM: PEPC is less sensitive to feedback inhibition by L-malate under light (C₄ plants) or at night (CAM plants) than in darkness (C₄) or during the day (CAM). While the view that …

In Vivo Regulatory Phosphorylation Site In C4-Leaf Phosphoenolpyruvate Carboxylase From Maize And Sorghum, Jin-An Jiao, Jean Vidal, Cristina Echevarría, Raymond Chollet

Department of Biochemistry: Faculty Publications

Reversible seryl-phosphorylation contributes to the light/dark regulation of C₄-leaf phosphoenolpyruvate carboxylase (PEPC) activity in vivo. The specific regulatory residue that, upon in vitro phosphorylation by a maize-leaf protein-serine kinase(s), leads to an increase in catalytic activity and a decrease in malatesensitivity of the target enzyme has been recently identified as Ser-15 in ³²P-phosphorylated/activated dark-form maize PEPC (J-A Jiao, R Chollet [1990] Arch Biochem Biophys 283: 300-305). In order to ascertain whether this N-terminal seryl residue is, indeed, the in vivo regulatory phosphorylation site, [³²P]phosphopeptides were isolated and purified from in vivo ³²P-labeled maize and …

Pharmacokinetics, Immune Response, And Biodistribution Of Iodine-131-Labeled Chimeric Mouse/Human Igg1, K 17-1a Monoclonal Antibody, Ruby Meredith, Albert Lobuglio, Walter Plott, Roger Orr, Ivan Brezovich, Charles Russell, Elizabeth Harvey, Michael Yester, Mansoor Saleh

Haematology and Oncology, East Africa

Pharmacokinetics,immunogenicity,and biodistributionof a 131I-Iabeledmouse/human chimenc monoclonal antibody (C17-1A) was studiedin six metastaticcoloncancerpatients. Pharmacokinetics obtalned from serum radioactivity or chi mera concentration were identical after 5 mCi of 131I(>17-1A withmeanalphahalf-livesof 17.6±2.3 and19.7±2.9 and meanbeta half-livesof 100.9±16.1 and 106.4±14.1 hr, respectively. HPLC analysis documented the monomeric chi rneiic17-lA withoutevidenceof immunecomplexesorfree 1311 None of the patients developed antibody after ‘31I-chi merle17-1A exposure.Radiolocalizationoccurredin known areas of disease >4 cm in all patients. The half-life of total body radioactivity was 58 ±7 hr by whole-body counts and 64 ±13 hr by urine measurements. Whole-body and bone marrow dose estimates ranged from 0.75-1 .03 and …

Phenology And Bionomics Of Culicoides (Diptera: Ceratopogonidae) In Glynn County, Georgia, Lisa M. Westberry

College of Graduate Studies: Theses & Dissertations (1964–2006)

Seasonal abundance and population densities of Culicoides species from sites in Glynn County, Georgia were surveyed using 15 New Jersey light traps during a 9-month period from April-December 1990. Environmental effects on the abundances were also analyzed. The three primary species present were: Culicoides furens (Poey), C. hollensis Mellander & Brues, and C. melleus (Coguillett). C. furens were collected year round, but predominantly during the summer. C. hollensis had a bimodal abundance (spring and fall) . C. melleus also had a bimodal abundance (late summer and fall).

Larval habitats of the primary species were identified …

A Temperature-Sensitive Mutant Of Escherichia Coli Affected In The Alpha Subunit Of Rna Polymerase, Majid Mehrpouyan

Electronic Theses and Dissertations

A temperature-sensitive mutant of Escherichia coli affected in the alpha subunit of RNA polymerase has been investigated. Gene mapping and complementation experiments placed the mutation to temperature-sensitivity within the alpha operon at 72 min on the bacterial chromosome. The rate of RNA synthesis in vivo and the accumulation of ribosomal RNA were significantly reduced in the mutant at 44$\sp\circ$C. The thermostability at 44$\sp\circ$C of the purified holoenzyme from mutant cells was about 20% of that of the normal enzyme. Assays with T7 DNA as a template showed that the fraction of active enzyme competent for transcription was reduced as a …