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Biomedical subjects

L Bogorad

Publications and source records attributed to L Bogorad.

At least 127 records · Page 7Linked to original sources

Light-induced, Dark-reversible Absorbance Changes in Roots, Other Organs, and Cell-free Preparations.

Irradiation of maize (Zea mays) roots and coleoptiles with visible light causes dark-reversible absorbance changes in these organs. There is an increase in absorbance near 440 nanometers and smaller increases below 410 nanometers and about 595 nanometers. Decreases in absorbance are observed at about 420 nanometers and minor ones at 537 and 575 nanometers. These responses are also observed in cell-free preparations from roots and coleoptiles if dithionite, NADPH, or NADH is added prior to illumination. The dose curve for these effects has a distinct maximum at 420 nanometers and a minor one at 575 nanometers. Difference spectra and dose response curves indicate that heme compounds such as cytochromes or, more probably, peroxidase complexes are the photoreceptive and chemical reacting molecules. Siroheme-containing proteins may also be taken into consideration.The light-induced absorbance changes have half-lives of more than 200 seconds and 100 seconds in roots of maize and soybean, respectively. Two reactions, each with first order kinetics, appear to be superimposed. The respective rate constants for maize roots are about 0.004 and 0.04 seconds(-1). The generation of the effect has a much shorter half-life dependent on light intensity and wavelength. Little deviations from first order kinetics were detected. Rate constants for corn roots range between 0.05 and 0.01 seconds(-1).Apart from the problem in which hemoproteins are involved, there is the problem of correlating the reaction of the photoreactive and chemically reacting molecules to macroscopic responses such as phototropism.

Journal Article↗

Purification and Characterization of RNA Polymerase from Fremyella diplosiphon.

We have purified and characterized a DNA-dependent RNA polymerase from the blue-green alga Fremyella diplosiphon. This enzyme, purified by gel filtration, DEAE-cellulose chromatography, and glycerol gradient centrifugation, is comprised of five polypeptide subunits. Their masses are 161,000, 134,000, 91,000, 72,000, and 41,000 daltons. Preparative electrophoresis of the purified enzyme on nondenaturing gels separates the 41,000-dalton polypeptide from the rest of the enzyme. The enzyme is extremely labile in the presence of a variety of salts of strong acids and bases; the purification procedure was devised to avoid exposure to such compounds.

Journal Article↗

Effects of erythromycin on membrane-bound chloroplast ribosomes from wild-type Chlamydomonas reinhardi and erythromycin-resistant mutants.

1. Treatment of wild-type cells of Chlamydomonas reinhardi with high concentrations of erythromycin results in increased recovery of membrane-bound chloroplast ribosomes, presumably by preventing polysomal runoff during harvesting of cells. No such membrane-retention effect is detected if erythromycin is added after harvesting of cultures, before cell breakage. 2. Growth of wild-type cells is inhibited by 10 microgram/ml erythromycin, but a concentration twice as high is required to increase recovery of membrane-bound wild-type ribosomes. On the other hand, the concentrations of erythromycin which inhibit growth of mutant ery-M1b produce a membrane-retention effect. Mutant ery-U1a is resistant to high concentrations of erythromycin and no membrane-retention effect is detectable at concentrations which produce one in wild type and ery-M1b. 3. These results can be reconciled by a two-point model of the mechanism of erythromycin action on chloroplast ribosomes in Chlamydomonas.

Chlamydomonas↗

Zea mays chloroplast ribosomal RNA genes are part of a 22,000 base pair inverted repeat.

Zea mays chloroplast rDNA exists in two identical units. Each unit contains one sequence for the 16, 23 and 5S rRNAs in the order given. The 16 and 23S sequences in each unit are separated by a 2100 base pair (bp) spacer. The DNA sequence for 5S RNA is closely linked to that for the 23S RNA. Within the above unit, the three RNAs are transcribed from a single DNA strand. The two rDNA units on the circular chloroplast DNA molecule are separated from each other by 18,500 bp in one direction and by 106,100 bp in the other direction. The two rDNA units have an inverted orientation with respect to each other. Each rDNA unit is part of a 22,000 bp sequence which is repeated with inverted orientation.

Base Sequence↗

Maize chloroplast DNA fragment encoding the large subunit of ribulosebisphosphate carboxylase.

In vitro linked transcription-translation of chloroplast DNA has been used to show that the large subunit of ribulose-1,5-bisphosphate carboxylase [3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] is encoded by Zea mays chloroplast DNA. A BamHI-generated chloroplast DNA sequence cloned in Escherichia coli is shown to direct the in vitro synthesis of this protein identified as large subunit by its size, serological properties, and limited proteolytic digestion products.

Journal Article↗

Action Spectra for Phycobiliprotein Synthesis in a Chromatically Adapting Cyanophyte, Fremyella diplosiphon.

The action spectra for phycocyanin production by the cyanophyte Fremyella diplosiphon shows maxima at 463 and 641 nm. The action spectrum for phycoerythrin production includes maxima at 387 and 550 nm. The maxima are based on a relative response rate well within the linear ascending portion of the dose response curves; the positions of the maxima are independent of the relative response rates chosen for reference over a 3-fold range although the comparative effectiveness of light at pairs of wavelengths varies with the standard used for comparison. These action spectra differ from those reported previously for Tolypothrix tenuis by Fujita and Hattori (Plant Cell Physiol. 3: 209-220) and by Diakoff and Scheibe (Plant Physiol. 51: 382-385) in that blue light strongly promotes phycobiliprotein synthesis in F. diphosiphon but has been reported to have little or no effect on T. tenuis.These data suggest that the photoreceptor or photoreceptors which regulate phycobiliprotein metabolism in F. diplosiphon may be either one or two metalloporphyrin complexes or double pigment systems consisting of a red or green light-absorbing photoreceptor which can be activated directly or by energy transferred from a blue light-absorbing receptor.

Journal Article↗

Hybridization of maize chloroplast DNA with transfer ribonucleic acids.

Hybridization of [125I] tRNA to chloroplast DNA indicates that 0.60-0.75% of maize chloroplast DNA contains sequences complementary to maize tRNA, corresponding to 20-26 tRNA cistrons. Green maize seedlings contain about twice the amount of chloroplast DNA-hybridizable tRNA as etiolated maize seedings. tRNA from green or etiolated maize seedlings was also aminoacylated in vitro with 21 labeled amino acids and then incubated with filters containing chloroplast DNA, tRNAs charging a total of at least 16 different amino acids hybridized with chloroplast DNA. Most of these plastid aminoacyl-tRNAs were present in higher concentrations in tRNA isolated from green maize seedlings, although there were several exceptions. The results are consistent with the hypothesis that a complete or nearly complete set or tRNAs can be transcribed from chloroplast DNA.

Amino Acids↗

Poly(adenylic acid)-containing RNA from plastids of maize.

Hybridization experiments with chloroplast DNA and 125I-labeled RNA from maize seedlings suggest that chloroplasts and etioplasts contain detectable amounts of RNA that contains poly(adenylic aicd) (poly (A)) and was transcribed from chloroplast DNA. About 6% of the total poly(A)-containing RNA isolated from maize seedlings hybridized to chloroplast DNA. Poly(A)-containing RNA could also be isolated directly from purified chloroplasts that were treated with ribonucleases to reduce cytoplasmic contamination. At least 65% of this poly(A)-containing RNA hybridized to chloroplast DNA. Chloroplast poly(A) tracts average about 45 nucleotides in length, one-half the average size of poly(A) tracts from whole cells. The poly (A) tracts themselves are probably added to plastid RNAs following their transcription, because maize chloroplast DNA was found not to contain poly(dT).

Centrifugation, Density Gradient↗

Light-induced increase in the activity of maize plastid DNA-dependent RNA polymerase.

Illumination of dark-grown maize plants induces a rapid rise in DNA-DEPENDENT RNA polymerase activity in plastids. Within 16 h illumination the enzyme activity is isolated plastids increases 3-4 fold as compared to other dark-grown plants illuminated for only 2 h. This change in activity is unaccompanied by either a quantitative rise in the amount of chloroplast RNA polymerase or qualitative changes in the purified enzyme. It is suggested that other factors may be present which interact either with the enzyme or the DNA template and that these factors are responsible for the light-induced enhancement of the RNA polymerase activity inplastids.

Chlorophyll↗

Endonuclease recognition sites mapped on Zea mays chloroplast DNA.

The closed-circular DNA molecules of 85 x 10(6) daltons from Zea mays chloroplasts were isolated, digested with the restriction endonucleases Sal I, Bam I, and EcoRI, and the resulting fragments sized by agarose gel electrophoresis. A map of maize chloroplast DNA showing the relative location of all the Sal I recognition sequences and many of the Bam I and EcoRI recognition sites was determined. A DNA sequence representing approximately 15% of the Zea mays chloroplast genome is repeated. The two copies of this sequence are in an inverted orientation with respect to one another and are separated by a nonhomologous sequence representing approximately 10% of the genome length. The inverted repeats contain the genes for chloroplast ribosomal RNAs.

Journal Article↗

Chloroplast membranes of the green alga Acetabularia mediterranea. II. Topography of the chloroplast membrane.

The localization of the chlorophyll-protein complexes inside the thylakoid membrane of Acetabularia mediterranea was determined by fractionating the chloroplast membrane with EDTA and Triton X-100, by using pronase treatment, and by labeling the surface-exposed proteins with 125I. The effects of the various treatments were established by electrophoresis of the solubilized membrane fractions and electron microscopy. After EDTA and pronase treatment, the membrane structure was still intact. Only the two chlorophyll-protein complexes of 67,000 and 152,000 daltons and an additional polypeptides were found in the membrane before the EDTA and pronase treatment. The 125,000 dalton complex seems to be buried inside the lipid layer. The 23,000 dalton subunit of the 67,000 dalton complex is largely exposed to the surface of the EDTA-insoluble membrane and only the chlorophyll-binding subunit of 21,500 daltons is buried inside the lipid layer.

Acetabularia↗

Chloroplast membranes of the green alga Acetabularia mediterranea. I. Isolation of the photosystem II.

1. In the presence of Triton X-100, chloroplast membranes of the green alga Acetabularia mediterranea were disrupted into two subchloroplast fragments which differed in buoyant density. Each of these fractions had distinct and unique complements of polypeptides, indicating an almost complete separation of the two fragments. 2. One of the two subchloroplast fractions was enriched in chlorophyll b. It exhibited Photosystem II activity, was highly fluorescent and was composed of particles of approx. 50 A diameter. 3. The light-harvesting chlorophyll-protein complex of the Photosystem II-active fraction had a molecular weight of 67 000 and contained two different subunits of 23 000 and 21 500. The molecular ratio of these two subunits was 2:1.

Acetabularia↗

Subunits of phycoerythrin from Fremyella diplosiphon: chemical and immunochemical characterization.

The alpha and beta subunits of the phycobiliprotein, phycoerythrin, isolated from the filamentous blue-green alga, Fremyella diplosiphon, have been separated by chromatography on Bio-Rex 70 ion exchange resin. Analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis shows no detectable cross-contamination of these subunit preparations. The molar extinction coefficients at 552 nm of the alpha and beta subunits in 8 M urea are 25,549 and 48,456, respectively. The amino acid compositions of the subunits are very similar. Molecular weights of the alpha and beta subunits are 19,500 and 21,700, respectively, based on the amino acid composition analyses. Antisera prepared against the alpha subunit reacts with the beta subunit, and vice versa. Tryptic peptide maps reveal that the subunits share share at least eight common tryptic peptides. These results indicate that the phycoerythrin subunits are chemically very similar.

Alkylation↗