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T Osumi

Publications and source records attributed to T Osumi.

At least 145 records · Page 8Linked to original sources
PubMed1980

Biosynthesis of ergosterol in cell-free system of yeast.

We previously proposed the occurrence of multiple pathways in the ergosterol biosynthesis of yeast, based on the results of examination of 14C-incorporation into sterols from L-[methyl-14C]methionine which was given to the intact cells of yeast. This led us to investigate the validity of the pathways by experiments with the cell-free system. Highly active cell-free extracts could be prepared by disruption of yeast cells with a Vibrogen Cell Mill in the presence of 0.1 mM dithiothreitol. This preparation catalyzed 14C-incorporation from [14C]methionine into ergosterol with a high yield. This preparation was found to be favorable for elucidation of ergosterol synthesis, since only a small amount of radioactivity was incorporated into fatty acid ester form of sterols which were reported to be inactive as a substrate for sterol synthesis reaction. Time course experiment of 14C-incorporation from [14C]methionine into various sterols under aerobic conditions showed that ergosta-8,24(28)-dien-3 beta-ol was a precursor for ergosta-7,24(28)-dien-3 beta-ol and that radioactivities were converted through ergosta-5,7,24(28)-trien-3 beta-ol and ergosta-5,7,22,24(28)-tetraen-3 beta-ol into ergosterol with time. In contrast, similar experiments under anaerobic conditions showed that ergosta-7,24(28)-dien-3 beta-ol accumulated and very little conversion of radioactivity into ergosterol occurred. In addition, the results indicated that oxygen was required for the introduction of double bond into 22 position as well as into 5 position. The results obtained with the cell-free system supported the validity of the proposal of multiple pathways of ergosterol synthesis in the intact cells.

Anaerobiosis↗
PubMed1979

Occurrence of two 3-hydroxyacyl-CoA dehydrogenases in rat liver.

3-Hydroxyacyl-CoA dehydrogenase was assayed for acetoacetyl pantetheine-reducing and acetoacetyl-CoA reducing activities in rat liver homegenates. Two isoenzymes of the enzyme, types I and II, were distinguished by the following procedures: trypsin treatment, heat treatment, CM-cellulose chromatography, antibody titration, and sucrose density gradient centrifugation of the light mitochondrial fraction. Type I enzyme was localized in mitochondria, and catalyzed the reduction of both acetoacetyl pantetheine and acetoacetyl-CoA. Type II enzyme was found mainly in peroxisomes, accompanied by a low activity in mitochondria or some other organelles, and was active with acetoacetyl-CoA but not with aceto acetylpantetheine. Both isozymes were induced by the administration to the rats of di-(2-ethylhexyl)phthalate, which enhances the peroxisomal beta-oxidation activity, but the extent of the induction of type II enzyme was much higher than that of type I enzyme. The activity of the former was found only in diethylhexylphthalate-treated rats.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1979

Studies on the delta 5-desaturation in ergosterol biosynthesis in yeast.

Studies were carried out on the delta 5-desaturation reaction in ergosterol biosynthesis with a particulate fraction of cell-free extract of yeast. A reduced pyridine nucleotide coenzyme and molecular oxygen were required for the reaction. It was shown that the enzyme activity is located in a fraction corresponding to microsomes. The reaction was inhibited by KCN, but not by CO. Menadione and potassium ferricyanide inhibited the NADPH- and NADH-dependent reactions, respectively, and cytochrome c inhibited both of them. These results suggested an involvement in delta 5-desaturation of a mixed function oxidase system resembling that for the fatty acyl-CoA desaturation reaction.

Ascorbic Acid↗
PubMed1978

Characterization of sterol-ester hydrolase in Saccharomyces cerevisiae.

A homgenate of Saccharomyces cerevisiae grown under semi-anaerobic as well as aerobic conditions was found to catalyze the hydrolysis of fatty acid esters of sterols in the presence of Triton X-100. The enzyme levels in cells grown under various conditions were similar and the enzyme had a broad substrate specificity for sterol esters. The enzyme was localized in the mitochondrial fraction for the aerobically grown cells and in the mitochondrial and cytosolic fractions for the semi-anaerobically grown cells.

Carboxylic Ester Hydrolases↗
PubMed1978

Ergosterol biosynthesis in yeast. Pathways in the late stages and their variation under various conditions.

[Methyl-14C]methionine was supplied to yeast cells under aerobic and anaerobic conditions for the investigation of the pathway for ergosterol biosynthesis after the methylation of the side-chain. Under aerobic conditions, the incorporation of radioactivity into ergosterol was high. With a limited oxygen supply, in contrast, the radioactivity was first accumulated in ergosta-7,24(28)-dien-3beta-ol and ergosta-8,24(28)-dien-3beta-ol, and then transferred to ergost-7-en-3beta-ol, ergost-8-en-3beta-ol and ergosta-7,22-dien-3beta-ol with time. Under strictly anaerobic conditions, a double bond was introduced neither to delta5 nor to delta22. The results of the tracer experiments suggested the operation of several pathways in the late stages of ergosterol biosynthesis. It was also suggested that the main pathways varied depending on the conditions such as oxygen supply and other factors. The above conclusion was supported by the results of the analyses of the sterol compositions of the cells grown under various conditions.

Aerobiosis↗
PubMed1978

Enhancement of fatty acyl-CoA oxidizing activity in rat liver peroxisomes by di-(i-ethylhexyl)phthalate.

The effect of di-(2-ethylhexyl)phthalate (DEHP) administration on cyanide-insensitive palmitoyl-CoA oxidizing activity in liver was studied. Two weeks of DEHP treatment increased the activity by one order of magnitude in male Wistar rats. A similar effect was also observed in male and female Sprague-Dawley rats and mice, but not in guinea pigs. When the liver was fractionated by differential centrifugation, the activity was concentrated in the light mitochondrial fraction. On the subfractionation of this fraction by sucrose density gradient centrifugation, the activity was distributed in a pattern similar to that of urate oxidase, but not resembling that of glutamate dehydrogenase. These data suggest that a fatty acyl-CoA oxidizing enzyme system which is located in peroxisomes is induced by the administration of DEHP.

Acyl Coenzyme A↗
PubMed1977

A novel pathway for L-citramalate synthesis in Rhodospirillum rubrum.

When [14C]propionate was incubated with a cell-free extract of Rhodospirillum rubrum in the presence of glyoxylate, ATP, CoA, Mg2+, and Mn2+, radioactivity was incorporated into mesaconate (MSA) as well as into beta-methylmalate (MMA) and citramalate (CMA). MSA was suggested to be an intermediate of the conversion of MMA to CMA based on the following observations. (i) When non-labeled MSA was added to the CMA-forming reaction system, radioactivity was trapped in MSA. (ii) When MSA was incubated with the cell-free extract, CMA was formed. (iii) The alpha-carboxyl group of CMA was shown to be derived from the beta-carboxyl group of MMA, implying that CMA was formed from MMA via MSA through successive dehydration and hydration. From the results of Sephadex G-10 column chromatography of the reaction products, beta-methylmalyl-CoA and mesaconyl-CoA were presumed to be involved in the reaction. A new CMA-forming metabolic pathway is proposed as follows: erythro-beta-methylamalyl-CoA leads to mesaconyl-CoA leads to MSA leads to L-CMA.

Glyoxylates↗
PubMed1976

[Pattern stimulation and salivary secretion (author's transl)].

Experiments were carried out to investigate the effect of temporal stimulus pattern of chorda or sympathetic stimulation on salivary secretion of the rabbit or the dog. Stimulation of short or long period was adopted. The latency and the velocity of secretion were recorded. The amount of secretion increased with the increase of the number of stimuli within moderate range. The optimum interval of chorda stimulation was about 50 msec, but it was somewhat longer for shorter period of stimulation. Various patterns of stimulation were adopted (1) by varying the interval, the number of stimuli and the period of stimulation, (2) by changing stepwise the interval halfway through stimulation and (3) by intermittent stimulation. Experiments on the effect of sympathetic stimulation were also reported. Fitness of Michaelis-Menten's equation for the relationship between the number of stimuli and the amount of secretion was tested. Moreover, the other equation that described this relation satisfactorily was derived from the hypothesis of superposition of unitary processes. It is suggested that the response of a excitable tissue to repetitive stimuli, if it be pattern sensitive, can not be predicted only from the intensity, the frequency and the number of stimuli without considering their temporal configuration.

Animals↗
PubMed1975

Formation of beta-methylmalate and its conversion to citramalate in Rhodospirillum rubrum.

Using a cell-free extract of Rhodospirillum rubrum, studies were made of the condensation reaction between propionyl-CoA and glyoxylate. When [14C]propionate was incubated with the extract in the presence of glyoxylate, ATP, CoA, Mg2+, and Mn2+, radioactivity was incorporated into several compounds. Two of the main products were characterized as citramalate (CMA) and erythro-beta-methylmalate (erythro-MMA) on the basis of their behavior compared with authentic samples of CMA and erythro-MMA in the following three analyses: (i) paper chromatography using two solvent systems, (ii) radio-gas chromatography on their methyl esters, and (iii) chemical conversion to readily crystallizable derivatives, that is, citramalyl chloralide for CMA, and thymine for MMA. The CMA was thought to be of L(+)-form based on the results of optical resolution with brucine and also its susceptibility to L(+)-citramalate lyase of Clostridium tetanomorphum. When the reaction was carried out with lower concentrations of the enzyme, only MMA was accumulated. However, when the reaction was allowed to proceed further after addition of higher concentrations of the enzyme and of excess semicarbazide to prevent further condensation, the amount of accumulated MMA was decreased and CMA was formed instead. Furthermore, the time course of MMA and CMA formation exhibited a pattern typical of a precursor-product relationship. From these results, it was concluded that MMA was formed by alpha-condensation between propionyl-CoA and glyoxylate, and that CMA was derived from MMA, possibly from its CoA derivative.

Adenosine Triphosphate↗
PubMed1975

[Pattern stimulation and action potential of frog skin (author's transl)].

Experiments were carried out to investigate the relationship between the pattern of stimulus of efferent or afferent nerve and the size of action potential of frog's skin. Two kinds of preparations were used: the excised nerve skin preparation and the spinal preparation. In the range of moderate intervals of stimuli, the size of skin action potential of the nerve-skin preparation became larger with the increase of number of stimuli, (i) during a definite period of stimulation with various frequencies, or (ii) during various long periods of stimulation with constant frequency. The size of potential depended not only on number of stimuli but also on their interval. The latter property may be called pattern-sensitive. In some state of preparation the size of potential evoked by a constant number of stimuli was nearly constant, being independent from stimulus frequencies, that is, pattern insensitive. Both kinds of results were obtained in cases of the spinal preparation. Michaelis-Menten's equation could be applied to describe the relation between the size of potential and the number of stimuli. However, there was no reason to interprete this relation with a simple concentration-effect relationship. Another hypothesis was then testoned on the supposition that the response is composed of sequentially superposed unitary processes each of which was promptly evoked by each stimulus and decayed exponentially. The calculated response curve reasonably fit with the observed data. Slower processes commonly called accomodation were excluded from the present discussion and only the effect of relatively short period of stimulation was examined.

Action Potentials↗