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Biomedical subjects

A Maeda

Publications and source records attributed to A Maeda.

At least 397 records · Page 22Linked to original sources

Isolation and characterization of defective interfering particle of Newcastle disease virus.

Newcastle disease virus grown in embryonated eggs was separated and purified by sucrose density gradient centrifugation into two distinct type of particles, B and T, the former being normal virus particles with high activities of hemagglutination, hemolysis, neuraminidase and infectivity, the latter being non-infectious virus particles with low activities of hemolysis and neuraminidase but high hemagglutination activity. B and T particles were shown to share a common antigen by immunodiffusion test. T particles were deficient in viral RNA, since they contained only 13s RNA in a small amount, whereas B particles possessed a large amount of 57s RNA and a small amount of 13s RNA. T particles interfered with the multiplication of normal Newcastle disease virus in primary cultures of chick embryo cells.

Animals↗

Induction of lesions in deoxyribonucleic acid by low molecular weight substances from normal and colicin E2-treated cells of Escherichia coli.

A fraction containing a variety of low molecular weight substances was extracted into 80% aqueous acetone from both a colicin E2-treated cell culture of Escherichia coli and an untreated one. The extract was divided into five fractions by Sephadex G15 chromatography. One of them, Fraction B, was separated into three subfractions by Sephadex G10 chromatography. Two subfractions, Fraction BI and Fraction BII, were further fractionated by several chromatographic systems. DNA was incubated with an aliquot from each of these fractions and was then analyzed by sedimentation in an alkaline sucrose density gradient. The activity which caused a decrease in the sedimentation coefficient of the DNA was found in some of these fractions. The activity from colicin E2-treated cells was compared with that from untreated ones. It was revealed that colicin E2 induces some increases in the activity toward DNA in one of the subfractions, Fraction BI, and also causes the appearance of a new species in another fraction, Fraction BII, which potentiates the activity in Fraction BI. These colicin E2-induced changes appeared at 5 min after the addition of colicin E2. The possible significance of such reactions for the action of colicin E2 are discussed.

Chromatography, Gel↗

Use of fluorescein mercuric acetate as a probe in studies of thiol-containing proteins.

The reaction of fluorescein mercuric acetate (FMA) with thiol groups is inhibited by barbital (Veronal). However, barbital has no effect on the reaction of 5,5'-dithiobis(2-nitrobenzoic acid) with thiol groups. Complex formation between FMA and barbital was shown by an increase in the fluorescence polarization of FMA in the presence of barbital. These findings indicate that the inhibitory effect of barbital is due to its interaction with FMA. The extent of this inhibition, which depends on the nature of the thiol-containing substance, presumably reflects the structure surrounding the thiol groups. Furthermore, the absorption and fluorescence properties of FMA conjugated with thiol groups are sensitive to the environment. Because of these properties, FMA can be used to investigate the environment of thiol groups.

Acetylcysteine↗

Isomeric composition of retinal chromophore in dark-adapted bacteriorhodopsin.

1. Retinal isomers extracted from the acid-hydrolysate of cetyltrimethylammonium bromide-treated dark-adapted bacteriorhodopsin (bRD) were analyzed in a high performance liquid chromatograph (HPLC) system. The extract from bRD contains almost equal molar amounts of both 13-cis retinal and all-trans retinal isomers. The extent of isomerization and the yield of both isomers during the isolation process were investigated by the application of the same extraction procedure to artificial bacteriorhodopsin reconstituted with 13-cis retinal isomer (13-cis bacteriorhodopsin) and also to light-adapted bacteriorhodopsin (bRL) which has been shown to contain only the all-trans isomer (all-trans bacteriorhodopsin). 2. A reconstituted bacteriorhodopsin, which had been prepared from apo-bacteriorhodopsin and an equimolar mixture of both 13-cis retinal and all-trans retinal isomers, showed an absorption spectrum having the same maximum wavelength as that of bRD even at the beginning of the reconstitution process. 3. Analysis of the photosteady states of bRD at -190 degrees C revealed that it was composed of two different species, one having 13-cis retinal and the other having all-trans retinal isomers in approximately equal molar amounts. These two also gave their respective photoproducts. 4. From these results it can be concluded that bRD contains both 13-cis retinal and all-trans retinal isomers in nearly equal molar amounts as its chromophore.

Bacteriorhodopsins↗

Interaction of a fluorescent reagent, fluorescein mercuric acetate, with nucleic acids.

Fluorescein mercuric acetate (fluorescein Hg Ac), which is a fluorescent thiol reagent, was shown to bind to various nucleic acids by measuring the changes in its absorption and fluorescence properties. Up to a critical concentration of free fluorescein Hg Ac (1-10(-7) M for calf thymus DNA, with 42% GC, and 2-10(-7) M for Micrococcus lysodeikticus DNA, with 72% GC) this reagent appears to bind selectively to single-stranded sections in DNA. Above this critical concentration, cooperative binding to double helical DNA occurs, and denatured DNA is obtained after removal of bound fluorescein Hg Ac by dialysis against 1 M KCl. These facts indicate that fluorescein Hg Ac causes the denaturation of double helical DNA prior to binding as has been shown in the case of methylmercuric hydroxide. The binding of fluorescein Hg Ac to DNA is much stronger than that of methylmercuric hydroxide. The number of total binding sites for fluorescein Hg Ac is close to the number of base pairs for both calf thymus DNA and M. lysodeikticus DNA. Furthermore, it was shown that fluorescein Hg Ac binds to thymidine, deoxyguanosine, poly(U) and poly(G). Since fluorescence quenching of fluorescein Hg Ac accompanies its complex formation with DNA and the affinity is markedly high as indicated by the association constant of 6.8-10(7) M(-1) for single-stranded calf thymus DNA, fluorescein Hg Ac can be used for the structural studies of small amounts of nucleic acids.

Binding Sites↗