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Biomedical subjects

W G Burton

Publications and source records attributed to W G Burton.

9 recordsLinked to original sources
PubMed1994

The active monomeric form of macrophage inflammatory protein-1 alpha interacts with high- and low-affinity classes of receptors on human hematopoietic cells.

Macrophage inflammatory protein-1 alpha (MIP-1 alpha) and its human homologue GOS19.1/LD78 are members of the C-C chemokine/intercrine family of secreted proteins. They have proinflammatory properties and also inhibit cell cycle progression of hematopoietic stem cells. Characterization of MIP-1 alpha receptor(s) has been confused because of its reported aggregation to inactive forms. Using a defined monomeric form of MIP-1 alpha that is biologically active for stem cell inhibition and induction of oxidative metabolism in polymorphonuclear cells, we report the detection of high- and low-affinity receptor classes on human leukemic CD34+ blast cells, promyelocytic cells, monocytes, peripheral blood neutrophils, and T cells. Both high- and low-affinity classes are expressed simultaneously in promyelocytes and neutrophils. The calculated kd for high-affinity receptors correlates with the concentrations of MIP-1 alpha required to induce a biologic effect on stem cells and neutrophils. Cross-linking studies show that MIP-1 alpha associates with two cell surface proteins with apparent molecular masses of 92 kD and 52 kD. Direct competition binding studies combined with studies on the inhibition of stem cells show that human and murine MIP-1 alpha have different receptor-binding and biologic properties.

Antigens, CD↗
PubMed1988

Separation of proteins by reversed-phase high-performance liquid chromatography. I. Optimizing the column.

In the process of developing a new analytical technology (the chromatophoresis process) which couples reversed-phase high-performance liquid chromatography (HPLC) to sodium dodecyl sulfate polyacrylamide gel electrophoresis in a real-time automated system, it was apparent that improvements in resolving power for the first-dimension (HPLC) separation were necessary. The present paper describes the optimization of the column for our initial work on reversed-phase HPLC separations. Polymeric (polystyrene) packings having particle diameters of 5 micron and pore diameters of 300 A were generally superior in terms of resolution, sample recovery and minimization of "ghosting". Optimum column dimensions were 50 x 1.0 mm I.D. for the flow-rates required in our system (10-100 microliter/min).

Chromatography, High Pressure Liquid↗
PubMed1988

Separation of proteins by reversed-phase high-performance liquid chromatography. II. Optimizing sample pretreatment and mobile phase conditions.

The effects of separation variables such as temperature, pH and composition of the mobile phase (including additives such as chaotropes, ion-pairing agents and surfactants), sample size and sample pretreatment for reversed-phase high-performance liquid chromatography (RP-HPLC) of proteins is examined. Experimental optimization of these parameters using the preferred instrumental and column conditions described previously lead to well behaved chromatographic performance for most proteins. This allowed us to achieve the required level of performance for the first dimension (RP-HPLC) separation of most protein samples by the chromatophoresis process.

Bacterial Proteins↗
PubMed1979

Messenger RNA of the large subunit of ribulose-1,5-bisphosphate carboxylase from Chlamydomonas reinhardi. Isolation and properties.

Polysomes specifically synthesizing the large subunit of ribulose-1,5-bisphosphate carboxylase were isolated from Chlamydomonas reinhardi cells by the indirect immunoprecipitation method. Electrophoretic analysis showed that the immunoprecipitated polysomes were of chloroplast origin. The mRNA coding for the large subunit which was purified from immunoprecipitated polysomes migrated at the 19-S position on sucrose density gradients, and its molecular weight was estimated to be 7.3 x 10(5) by acid-urea/agarose gel electrophoresis. The mRNA was translated in vivo with a cell-free protein-synthesizing system derived from Escherichia coli to give full-length large-subunit polypeptides.

Carboxy-Lyases↗
PubMed1979

Role of methylation in the modification and restriction of chloroplast DNA in Chlamydomonas.

The different metabolic paths followed by homologous chloroplast DNAs of maternal and paternal origins in zygotes of Chlamydomonas were examined by prelabeling parental cells, before mating them, with [3H]adenine, [3H]thymidine, and [3H]deoxycytidine. Within 6 hr after mating, maternal chloroplast DNA was extensively methylated to 5-methylcytosine and its bouyant density decreased. Paternal chloroplast DNA was largely degraded. Some radioactivity from deoxycytidine of maternal origin reappeared in thymine, and residual paternal DNA contained radioactivity in a base tentatively identified as uracil. These results confirm and extend our previous findings and support our hypothesis that modification (methylation) and restriction enzymes determine maternal inheritance of chloroplast DNA and that the two parental DNAs have different metabolic fates within the zygote.

Adenine↗
PubMed1977

A site-specific single-strand endonuclease from the eukaryote Chlamydomonas.

We have found a unique deoxyribonuclease in extracts of the eukaryotic green alga Chlamydomonas. When incubated with viral DNA from adenovirus-2, this enzyme produces discrete fragments that form bands upon electrophoresis in an agarose gel. Site specificity of the enzymatic cleavage examined by identifying the 5'-terminal nucleotides in cleaved adenovirus-2 DNA and by studies with synthetic polynucleotides of defined sequence, indicates that the initial endonucleolytic cleavage occurs at a site containing a deoxythymidine residue. Electron microscopy of cleaved adenovirus-2 DNA revealed single-strand segments within duplex DNA. We propose that the enzyme acts by making initial site-specific single-strand incisions, followed by subsequent excision on the same strand, producing a gapped duplex molecule; and that double-strand scissions result from limited occurrence of overlapping single-strand gaps on complementary strands.

Base Sequence↗
PubMed1973

Can a non-Mendelian mutation affect both chloroplast and mithchondrial ribosomes?

Chloroplast ribosomes isolated from a spectinomycin-resistant mutant (spr-1-27-3) of Chlamydomonas reinhardtii that displays non-Mendelian inheritance fail to bind labeled antibiotic, in contrast to ribosomes from wild-type cells. In vitro resistance of this mutant appears to result from the absence of a specific protein in the small subunit of the chloroplast ribosome. However, chloroplast protein synthesis in the mutant and wild type shows identical sensitivity to spectinomycin in short-term in vivo experiments where ribulosediphosphate carboxylase serves as the marker. Long-term experiments demonstrate that the mutant can grow in the presence of spectinomycin only when acetate is supplied as a carbon source. Mitochondrial structure and function of the mutant are not affected by the antibiotic, whereas chloroplast structure and function are. Apparently, the mitochondrion, rather than the chloroplast, of this mutant is resistant to spectinomycin in vivo. We hypothesize that the gene product of the spr locus is a protein common to both chloroplast and mitochondrial ribosomes. The mutant gene product, in vivo, confers resistance on mitochondrial, but not chloroplast, ribosomes. We suppose that the mutant spr protein loosely attaches to chloroplast ribosomes in vivo so that the antibiotic is bound and blocks protein synthesis, but it dissociates during isolation, resulting in loss of the binding site.

Binding Sites↗