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Articles 1 - 13 of 13

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

Analysis And Characterization Of The Swine Major Histocompatibility Complex , Wen-Rong Lie Jan 1987

Analysis And Characterization Of The Swine Major Histocompatibility Complex , Wen-Rong Lie

Retrospective Theses and Dissertations

Using SLA inbred and recombinant miniature swine as models, this dissertation describes: (1) Quantitative differences in GLO enzyme activity associated with the SLA complex; (2) The mapping of swine C2, Bf, and C4 genes to the SLA complex; and (3) The preparation and characterization of MAb to swine leukocyte antigens. In the first project, no electrophoretic differences in GLO among miniature swine were detected. However, GLO enzyme activity in SLA[superscript] a/a cells was consistently higher than in SLA[superscript] d/d cells, whereas GLO activity of SLA[superscript] a/d heterozygotes was intermediate. Using this information, a family ...


Physical Studies Of Adenylosuccinate Synthetase , Michael Brian Bass Jan 1987

Physical Studies Of Adenylosuccinate Synthetase , Michael Brian Bass

Retrospective Theses and Dissertations

To determine the chemical mechanism of the reaction catalyzed by adenylosuccinate synthetase, positional isotope exchange studies were performed. Positional isotope exchange from the (beta)-(gamma) bridge to the (beta) nonbridge position of (gamma)-('18)O GTP was followed using ('31)P NMR. The positional isotope exchange was found to occur in the presence of either IMP or IMP and succinate. The exchange did not occur in the presence of asparate. These results support a reaction mechanism which involves formation of a 6-phosphoryl-IMP intermediate with subsequent attack by aspartate to form adenylosuccinate as originally proposed by Lieberman in 1956 Lieberman, I ...


Determination Of The Reactivity Of Cytotoxic Immune Cells With Preimplantation Mouse Embryos , Mark A. Ewoldsen Jan 1987

Determination Of The Reactivity Of Cytotoxic Immune Cells With Preimplantation Mouse Embryos , Mark A. Ewoldsen

Retrospective Theses and Dissertations

Cytotoxic immune cells were used in an assay, MELIA (mixed embryo leukocyte interaction assay) to test the ability of the cells to kill blastocyst stage embryos. The cytotoxic immune cells generated for use in this study, cytotoxic T lymphocytes (CTLs), natural killer (NK) cells, and lymphokine activated killer (LAK) cells were shown to have phenotypic and cytolytic characteristics similar to those reported by other investigators. The lysis of the blastocysts in the MELIA was determined by measuring the inhibition of blastocoel retention and/or by the inhibition of incorporation of tritiated thymidine (('3)H-TdR) into embryonic DNA. Blastocysts which possess ...


Effects Of Clofibric Acid On Porcine Hepatocytes: A New Model For The Study Of Peroxisomal Metabolism , Kenneth Wayne Turteltaub Jan 1987

Effects Of Clofibric Acid On Porcine Hepatocytes: A New Model For The Study Of Peroxisomal Metabolism , Kenneth Wayne Turteltaub

Retrospective Theses and Dissertations

Data on interspecies peroxisomal metabolism are limited almost entirely to rodents. Species differences are likely and the application of rodent data to man is questionable. An alternative to rodent models is desirable. Porcine tissue offers such an alternative and was explored. Porcine hepatocyte organelles were separated by isopycnic sucrose gradient centrifugation from livers of 6-month-old Yorkshire pigs. The presence of a peroxisomal palmityl-CoA oxidizing pathway, peroxisomal superoxide dismutase (SOD), and a peroxisomal NAD:aldehyde dehydrogenase (ALDH) with high K[subscript] m for acetaldehyde was demonstrated. Peroxisomal palmitate oxidizing capacity was found to be equal to that of the surviving mitochondria ...


The Mouse Major Histocompatibility Complex And Age-Related Changes In The Murine Immune System , Terry Euclaire Meyer Jan 1987

The Mouse Major Histocompatibility Complex And Age-Related Changes In The Murine Immune System , Terry Euclaire Meyer

Retrospective Theses and Dissertations

A and C57BL/6 inbred mice were used as models for investigating interactions between aging processes and the genes that regulate the immune system, the major histocompatibility complex (MHC, H-2) genes. The C57BL/6 (H-2[superscript] b) mice exhibited significantly longer mean lifespans than the A (H-2[superscript] a) mice, as has been observed by other investigators. Backcross and F[subscript] 2 generation offspring from the parental A and C57BL/6 mice were tested for dependence of longevity upon H-2 type. The effects of H-2 type upon lifespan were statistically significant only for the oldest surviving (i.e., "10[superscript ...


A Novel Insulin From The Bullfrog: Its Structure And Function In Protein Secretion By Hepatocytes , Jocelyn Jean Hulsebus Jan 1987

A Novel Insulin From The Bullfrog: Its Structure And Function In Protein Secretion By Hepatocytes , Jocelyn Jean Hulsebus

Retrospective Theses and Dissertations

Bullfrog insulin was extracted and purified from the pancreas of Rana catesbeiana adults using gel filtration and reverse phase high performance liquid chromatography. Amino acid analysis of bullfrog insulin revealed 52 amino acids instead of the most common number of 51. Hagfish insulin is the only other known insulin with 52 amino acids. The most unique feature of bullfrog insulin is a two amino acid extension on the amino terminus (A1) of the A chain. This is the only insulin to date that has an extension at this position. In mammals, the deletion of A1 glycine or addition of a ...


The Construction And Properties Of An Integrable Plasmid For Genomic Analysis In Staphylococcus Aureus , John Bernard Luchansky Jan 1987

The Construction And Properties Of An Integrable Plasmid For Genomic Analysis In Staphylococcus Aureus , John Bernard Luchansky

Retrospective Theses and Dissertations

An improved system for transforming protoplasts of Staphylococcus aureus with plasmid DNA was developed to facilitate the construction of an integrable plasmid. Protoplast transformants were recovered at higher frequencies when the protoplasts were warmed to 56 C immediately prior to the addition of cesium chloride-purified plasmid DNAs. More transformants were recovered when the protoplast-DNA mixture was plated on R agar than on DM3 medium, and when the selective agent was incorporated directly in the medium rather than supplied in an agar overlay. More transformants were recovered when the protoplast-DNA mixture was incubated at 30 C for 3.5 h before ...


The Metabolism And Localization Of Two Proteins During Heat Shock Of Tetrahymena Thermophila , Thomas William Mcmullin Jan 1987

The Metabolism And Localization Of Two Proteins During Heat Shock Of Tetrahymena Thermophila , Thomas William Mcmullin

Retrospective Theses and Dissertations

When T. thermophila is subjected to hyperthermal stress, it immediately but transiently represses the synthesis of the normal array of proteins (non-hsps) and induces the synthesis of the heat shock protein (hsps). These changes in protein synthetic patterns are controlled at both the transcriptional and translational levels;To determine whether the translational regulatory events could be correlated with changes in the protein synthetic machinery, heat shock induced modifications of the ribosomal proteins were investigated. Of particular interest was the heat shock induced association of a 22 kDa protein, p22, with the small subunit of the ribosome. The synthesis of p22 ...


Regulation Of Glycogen Phosphorylase By Its Amino-Terminal Region , William Richard Harris Jan 1987

Regulation Of Glycogen Phosphorylase By Its Amino-Terminal Region , William Richard Harris

Retrospective Theses and Dissertations

In the first section of this dissertation, the role of the amino-terminal region of glycogen phosphorylase was studied by removal of this region and determination of the properties of the truncated phosphorylase analog. Limited proteolysis of phosphorylase by subtilisin Carlsberg in the presence of the phosphorylase inhibitor caffeine produced a phosphorylase analog, named phosphorylase b[subscript]sp S', missing only the amino-terminal 16 to 18 residues. Phosphorylase b[subscript]sp S' was purified by anion-exchange high performance liquid chromatography (HPLC). The maximal velocity of phosphorylase b[subscript]sp S' was 47% higher than that of phosphorylase b, while the Michaelis ...


Mechanistic Aspects Of The Low-Molecular-Weight Phosphatase Activity Of Calcineurin: A Model For Phosphotyrosyl-Protein Phosphatases , Bruce L. Martin Jan 1987

Mechanistic Aspects Of The Low-Molecular-Weight Phosphatase Activity Of Calcineurin: A Model For Phosphotyrosyl-Protein Phosphatases , Bruce L. Martin

Retrospective Theses and Dissertations

The low-molecular-weight phosphatase activity of the calmodulin-activated protein phosphatase, calcineurin, has been investigated as a model reaction for the dephosphorylation of phosphotyrosyl residues. The substrate specificity, kinetic mechanism, and chemical mechanism have been studied via kinetic means. Calcineurin is found to dephosphorylate aryl phosphate esters with a nonconstant maximal velocity, in contrast to the situation observed with the nonspecific acid and alkaline phosphatases from various sources. Product inhibition kinetics demonstrated that the kinetic mechanism of calcineurin is a random uni-bi. Calcineurin failed to show phosphotransferase activity with two acceptors functional with the nonspecific acid and alkaline phosphatases. The simplest model ...


Studies On The Transacylation Of Retinol Catalyzed By Acyl Coenzyme A:Retinol O-Acyltransferase , Mark David Ball Jan 1987

Studies On The Transacylation Of Retinol Catalyzed By Acyl Coenzyme A:Retinol O-Acyltransferase , Mark David Ball

Retrospective Theses and Dissertations

Acyl coenzyme A:retinol O-acyltransferase (ARAT), a microsomal enzyme, is thought to catalyze the transacylation reaction whereby retinol is esterified in vivo. Therefore, by means of an in-vitro assay method measuring net synthesis of retinyl esters, properties of the enzyme in various tissues were studied;ARAT was characterized in the liver of a polar bear, a species with exceptionally large vitamin-A reserves. The activity observed was unusually high. Vitamin A was present in the liver at 8050 [mu]g/g tissue, with 98% of the vitamin in its ester form. However, the activity of ARAT in rat mammary tumor, a ...


Isolation And Properties Of The Catalytically Active [Gamma] Subunit Of Phosphorylase Kinase , Scott Martin Kee Jan 1987

Isolation And Properties Of The Catalytically Active [Gamma] Subunit Of Phosphorylase Kinase , Scott Martin Kee

Retrospective Theses and Dissertations

Phosphorylase kinase is a complex enzyme in both structure and regulation. It has a subunit stoichiometry of ((alpha)(beta)(gamma)(delta))(,4), and the (alpha), (beta), and (delta) subunits have known regulatory functions, whereas the (gamma) subunit has a catalytic site. The (gamma) subunit, separated from the holoenzyme by reversed-phase high-performance liquid chromatography (HPLC), had no activity in the usual assay mixture, but was reactivated in the presence of Ca('+2) and calmodulin (which is identical to the (delta) subunit). The renaturation and reactivation process, which is totally dependent on Ca('+2) and calmodulin, is optimized under specific conditions of ...


11-Cis Retinoids: Methods Of Analysis And Handling, And Regeneration In Excised Bovine Eyes , Gary Marshall Landers Jan 1987

11-Cis Retinoids: Methods Of Analysis And Handling, And Regeneration In Excised Bovine Eyes , Gary Marshall Landers

Retrospective Theses and Dissertations

A statistical optimization technique, based on estimating a three-dimensional response surface for resolution over a range of mobile-phase compositions, was applied to the separation of isomers of retinol in normal-phase HPLC (high-performance liquid chromatography). The optimum solvent mixture for resolution of the 11-cis from the 13-cis isomer was very different from the optimum mixture for resolution of the 9-cis from the all-trans isomer. The effect of solvent selectivity on resolution was found to be much greater for resolution of different retinoid classes than for resolution of isomers within a class;No isomerization of all-trans retinal, retinyl palmitate or retinol in ...