Life Sciences Commons^™

A Characterization Of Extractable, Hydroxylated Fatty Acid Bearing Components In Legionella Pneumophila, Jonathan R. Lane

Electronic Theses and Dissertations

Extraction of the lipids of Legionella pneumophila yields phases unlike those produced from other Gram-negative bacteria. A viscous interface forms between the aqueous (wash) and organic phases. More than half of the hydroxylated fatty acids were found distributed between the aqueous phase and the interfacial material, fractions in which such constituents have not been reported in other Gram-negative species. It was further observed that after the material from the aqueous/interfacial phase was dissolved in methanol or chloroform/methanol (2:1 (V/V)), the addition of acetone would create a white, flocculent precipitate. Analyses showed that the supernatant contained fatty acids that were nonhydroxylated …

Probing Protein-Protein Interactions Among Proteins Of A Nonaggregated Fatty Acid Synthetase From Euglena Gracilis Variety Bacillaris, Sande G. Williams

Electronic Theses and Dissertations

Enoyl-acyl carrier protein (ACP) reductase from chloroplast nonaggregated fatty acid synthetase (FAS) of Euglena gracilis variety bacillaris was purified to a single band on a denaturing polyacrylamide gel. The enzyme was partially characterized with respect to substrate specificity, reduced nucleotide requirement, and the effect of ACP and Ca$\sp{++}$ on enzyme activity. Antibodies against the purified protein were raised in hens and isolated from eggs. ACP was purified from Euglena in yields of about 1mg/100g (wet weight) of cells. Antibodies were raised against the purified protein. ACP antibodies inhibited the Euglena chloroplast FAS using Euglena or E. coli ACP as a …

Human Lung Mast Cell Tryptase Isozymes: Separation And Examination Of Structural And Functional Differences, Susan S. Little

Electronic Theses and Dissertations

Tryptases are trypsin-like enzymes found in mast cell granules. Although in vivo substrates have not been positively identified, tryptases cleave a limited number of potential physiological substrates in vitro, including high molecular weight kininogen (HMWK) and vasoactive intestinal peptide (VIP). Purified human lung mast cell tryptase (HLT) apparently exists as a tetramer with an M$\sb{\rm r}$ of 135-144 kDa by gel filtration, whereas SDS-PAGE yielded two bands of M$\sb{\rm r}$ 29 Kda and 33 Kda. Tryptases are resistant to inhibition by most natural trypsin inhibitors and display some affinity for heparin. The existence of tryptase isozymes has been implied from …

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 …

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 …

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 …

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 …

Purification And Characterization Of Enoyl-Acp Reductase From Euglena Gracilis, Margie M. Tucker

Electronic Theses and Dissertations

Enoyl-(acyl-carrier-protein) reductase was purified from the phytoflagellate Euglena gracilis. Its purification employed DEAE-Sephacel chromatography, Matrex Orange chromatography, and affinity chromatography using acyl carrier protein (ACP) covalently bound to Sepharose as the affinity ligand. Matrex Orange chromatography resolved two different enoyl-ACP reductases having different characteristics. Euglena gracilis appears to resemble higher plants in the possession of two isoforms of this enzyme. Antibodies specific for the cofactor binding site of NADP (H)-requiring dehydrogenases were obtained. They were isolated from a polyclonal population of antibodies directed against yeast glucose-6-phosphate dehydrogenase by affinity chromatography using chicken liver malic enzyme as the affinity ligand. The …

Variants And Polymorphisms Of Three Repetitive Dna Families In The Human Genome, Robert M. Roudabush

Electronic Theses and Dissertations

A novel 0.6 kb LINE family in human DNA, designated L2Hs, has been described (Musich and Dykes 1986). Studies employing clone N6.4, containing three 0.6 kb segments of this family, indicate that these sequences are interspersed and moderately repetitive. Two additional variant sequences of the L2Hs family, N6.1 and N6.3, have been identified. Restriction mapping of each cloned segment indicates similarities among N6.4, N6.3 and N6.1. When the cloned DNAs were cleaved with restriction enzymes and subjected to cross-hybridization, each cloned insert produced a pattern indicating that the sequences contained in N6.1 and N6.3 are represented in at least one …

The Bacterial Cell Membrane: Berlin Wall Of The Cell, Hans Kornberg

DSLS 1987-1988

(This information was taken from the Distinguished Scientist Lecture Series Program 1983-1984).

RECORDING UNAVAILABLE.

Born in Herford, Germany, Dr. Kornberg is Sir William Dunn Professor of Biochemistry, University of Cambridge, and Master of Christ's College, Cambridge. Dr. Kornberg received the B.Sc. and Ph.D. degrees from the University of Sheffield, the M.A. and D.Sc. degrees from the University of Oxford, and the Sc.D. degree from the University of Cambridge. He did postdoctoral research at Yale University, the University of California, Berkeley, and the Public Health Research Institute (New York City) before returning to England to join the Medical Research Council's Cell …

Purification And Characterization Of An Inhibitor Of Thymidine Uptake From Culture Supernatants Of Human Tonsil Lymphocytes, Abdolreza Zarnegar

Electronic Theses and Dissertations

Lymphocytes from human tonsils were cultured in the absence of serum for 3 days. In the presence of the concentrated culture supernatant the proliferative response of PBL, to con A, as measured by the uptake of ('3)H-tdr, was significantly reduced. The suppressor substance was referred to as SMAL (suppressor of mitogen activated lymphocytes). The estimated molecular weight of SMAL under nondenaturing conditions was 100,000-300,000. SMAL also suppressed the incorporation of ('3)H-tdr by a variety of mouse and human tumor cell lines. The activity of SMAL was sensitive to pronase and heating at 100(DEGREES)C for 30 minutes but insensitive to RNase. …

Human Lung Tryptase Purification And Characterization, Timothy J. Smith

Electronic Theses and Dissertations

Human lung tryptase (HLT), a mast cell derived trypsin-like enzyme, was isolated from whole human lung tissue obtained at autopsy. Increased yields from this purification process allowed extensive characterization of the enzyme. One of the critical steps in the purification scheme was the use of a linear heparin gradient to elute active material from cellulose phosphate. Gel filtration studies in 1.0 M NaCl yielded an apparent M(,r) of 135,000, and subsequent electrophoresis on sodium dodecyl sulfate-polyacrylamide gels demonstrated the presence of two active species with apparent M(,r) = 30,900 and 31,600. Enzymatic activity was sensitive to NaCl concentrations above 0.05 …

Effect Of Fluoroacetate On Amino Acid Metabolism / W. J. Nicklas, D. D. Clarke, And S. Berl. Chem. Dept., Fordham Univ., And Col. Of Physicians And Surgeons, Columbia Univ., New York, N. Y., William J. Nicklas, Donald Dudley Clarke Phd, Soll Berl

Chemistry Faculty Publications

Guinea pig brain cortex slices incubated in media containing U-14-aspartate or glutamate (GA) (J. Neurochem. in press) form glutamine ( GM) of relative specific activity (RSA)> 1 (GA=l). In the presence of 10-3 fluoroacetate (FA) the RSA of GM decreased to values considerably < 1 without any changes in levels of amjno acids; the % of label in GA doubled whilA that in GM decreased correspondingly. Similar results were obtained with 1-14 C-acetate as tracer in vivo (mice) and in vitro. RSA' s of Grvl > 1 have been explained by the compartmentation of GA in cerebral cortex. Consequently the effect of GA may be explained by an inhibition of the flow of metabolites through that pool of GA used preferentially for GM synthesis. This is consistent with the concept of the existence, in brain, of more than one citric acid cycle, one …

Saponin Content And Some Pod And Blossom Characteristics Of Alfalfa As Related To Seed Infestation By The Alfalfa Seed Chalcid, Ronald D. Morse

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The alfalfa seed chalcid, Bruchophagus ruddi Guss., is a jet-black hymenopteran wasp. The destructive nature of this pest has been recognized since the latter part of the nineteenth century. Every year thousands of acres of alfalfa seed are destroyed, with infestation reaching as high as 8 5 percent in some areas. In Utah the chalcid annually ruins from 5 to 25 percent of the alfalfa seed. Much of this damage goes unnoticed, as infested seed is commonly blown out in the trash during harvesting and cleaning operations . The extent of damage is not restricted to the United States. Wherever …