PubMed Health⌕ Search

Biomedical subjects

S Omata

Publications and source records attributed to S Omata.

At least 19 recordsLinked to original sources
PubMed1992

Isolation and some properties of a 34-kDa-membrane protein that may be responsible for ribosome binding in rat liver rough microsomes.

We have isolated, by hydroxyapatite chromatography with a non ionic detergent and a high salt concentration, a non-glycosylated, membrane protein with a relative molecular weight of 34 kDa that had previously been found to be a major constituent of the membrane protein fraction showing ribosome-binding activity derived from rat liver rough microsomes (RM). The isolated 34 kDa protein (p34), when incorporated into a liposome model membrane, exhibited significant binding activity toward ribosomes, its binding properties being similar to those observed with intact RM. Immunochemical analyses using antibodies directed against p34 suggested that it is a membrane-embedded RM surface protein, which is specifically localized in ribosome-attached organelles and widely distributed among mammalian tissues. These results would constitute evidence that p34 is a likely candidate for an RM ribosome-binding protein.

Animals↗
PubMed1992

In vitro formation of estrogen receptor-heat shock protein 90 complexes.

We previously showed that the 9 S estrogen receptor can be reconstituted from purified vero ER (estradiol binding subunit) and purified hsp 90 (heat shock protein 90) in vitro [Inano, K. et al. (1990) FEBS Lett. 267, 157-159]. In this study, we further characterized our reconstitution system to investigate the mechanism underlying the formation of 9 S ER. When a vero ER preparation stored at 4 degrees C for more than 20 h after affinity chromatography was used for the reconstitution of 9 S ER, 0.5 M NaSCN was essential, but not Na2MoO4 or other reagents. When, however, vero ER was used within 3 h after dissociation from an affinity resin, 9 S ER could be reconstituted in a relatively high yield without NaSCN. Moreover, if such a fresh vero ER preparation was used, 9 S ER could be reconstituted in the absence of NaSCN from not only unoccupied vero ER but also the occupied form. From these results it was suggested that the conformation of purified vero ER tends to change quickly in a time dependent manner, and so a chemical perturbant, NaSCN, is generally necessary for the reconstitution of 9 S ER from purified vero ER and purified hsp 90. The concentration of hsp 90 required for the reconstitution was only about 1.0 microM, which was lower than its physiological concentration. Based on these results, the mechanism underlying the formation of 9 S ER was discussed.

Animals↗
PubMed1989

Differential effects of methylmercury on the synthesis of protein species in dorsal root ganglia of the rat.

Dorsal root ganglia from control and methylmercury (MeHg)-treated rats were incubated in vitro with 35S-methionine ant the proteins synthesized were analyzed by two-dimensional electrophoresis. The double labelling method, in which proteins of control dorsal root ganglia labelled in vitro with 3H-leucine were added to each of the two samples as an internal standard, was used to minimize unavoidable errors arising from the resolving procedure itself. The results obtained showed that the effect of MeHg on the synthesis of proteins in dorsal root ganglia was not uniform for individual protein species in the latent period of MeHg intoxication. Among 200 protein species investigated, 157 showed inhibition of synthesis close to that of the total proteins in the tissue (68% of the control). Among the remaining protein species, 20 showed real stimulation of synthesis, whereas 7 were moderately inhibited and 16 were inhibited more strongly than the total proteins in the tissue. These results suggest that the effect of MeHg on the synthetic rates for protein species in dorsal root ganglia differs with the species, and that unusual elevation or reduction of the synthesis of some protein species caused by MeHg may lead to impairment of normal nerve functions.

Animals↗
PubMed1988

In vivo and in vitro effects of methylmercury on the activities of aminoacyl-tRNA synthetases in rat brain.

The activities of six aminoacyl-tRNA synthetase species were determined using enzyme preparations partially purified from the brains of control and methylmercury (MeHg)-treated rats. The activities of Asp-, Leu- and Tyr-tRNA synthetases were significantly reduced in the brains of MeHg-intoxicated rats, whereas those of Lys- and Met-tRNA synthetases remained unchanged. In contrast, the activity of His-tRNA synthetase was significantly increased in the symptomatic phase of MeHg intoxication. The activities of these six aminoacyl-tRNA synthetases in the control brains were affected to different extents on the direct addition of MeHg to the assay system in vitro. No positive correlation was observed between the in vivo and in vitro effects of MeHg on the enzyme activities. These results indicate that the aminoacylation of tRNA is one of the actions of MeHg, which leads to inhibition of protein synthesis, and it is suggested that the syntheses of cellular proteins may be modified in different ways by MeHg, depending on their amino acid compositions.

Amino Acyl-tRNA Synthetases↗
PubMed1987

Decrease in protein phosphorylation in central and peripheral nervous tissues of methylmercury-treated rat.

The protein phosphorylation in extracts of nervous tissues of rats acutely exposed to methylmercury chloride (seven daily injections of 10 mg methylmercury chloride/kg body weight) was examined. In the brain, the phosphorylating activity was dependent on cAMP and Mg2+. The effect of methylmercury on the phosphorylation of brain proteins, including tubulin and MAP-2, was hardly discernible. In peripheral nervous tissues such as the dorsal and ventral roots, sciatic nerves and dorsal root ganglia, the phosphorylating activity was dependent on Ca2+, and the maximal activity was obtained when the tissues were extracted in the presence of 1% Triton X-100. SDS-Polyacrylamide gel electrophoresis revealed that the major phosphorylated proteins in the peripheral tissues were myelin proteins. The effects of methylmercury were not uniform regarding protein species and tissues. The most marked changes were observed in sciatic nerves, in which phosphorylation of the 33 kDa, 28 kDa, 19 kDa, 18 kDa and 15 kDa proteins was significantly decreased in the symptomatic phase of intoxication.

Animals↗
PubMed1986

Species difference between rat and hamster in tissue accumulation of mercury after administration of methylmercury.

The accumulation of mercury in tissues of the rat and hamster was determined after the administration of a single dose of 203Hg-methylmercury chloride (10 mg/kg body weight). On day 2, the mercury contents of hamster tissues were higher than those of rat tissues, except for red blood cells, in which the mercury content was about 6-fold higher in the rat than in the hamster. After that time, the mercury content of hamster tissues decreased rather steeply and on day 16 it had reached 14-25% in nervous tissues and 7-15% in other tissues, of the levels on day 2. In the rat, on the other hand, the mercury content of nervous tissues on day 16 was higher than that on day 2 (106-220%), except for dorsal roots and dorsal root ganglia, which showed slight decreases (75-94% of the levels on day 2). In non-neural tissues, the decreases up to day 16 were also small (71-92% of the levels on day 2). Thus, both the uptake and elimination of mercury seem to be more rapid in the tissues of hamster compared with those of the rat. Similar trends of mercury accumulation and elimination were observed when animals received multiple injections of methylmercury that induced acute methylmercury intoxication. Significant biotransformation of the injected methylmercury to inorganic mercury was detected in the liver, kidney and spleen of both animal species.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Alteration in RNA synthesis in the dorsal root ganglia of methylmercury-treated rats.

The effect of methylmercury in vivo on the incorporation of 3H-uridine in vitro into RNA of dorsal root ganglia of the rat was examined. Modified methods for the incubation of the tissue and extraction of RNA were applied to adequately determine the rate of RNA synthesis. Methylmercury significantly decreased the RNA content and RNA synthetic activity only in the symptomatic period, while uptake of the precursor into the acid-soluble pool remained unchanged. These results indicate that the previously reported inhibition of protein synthesis in dorsal root ganglia at an early phase of methylmercury intoxication was not due to impairment RNA synthesis in this tissue.

Animals↗
PubMed1985

Properties of rat liver signal peptidase reconstituted into liposomes.

EDTA/KCl- or pyrophosphate-treated rough microsomes of rat liver clearly showed the co-translational cleavage of pre-human placental lactogen and translocation of the product into membrane vesicles. The signal peptidase fraction was isolated by chromatography on Sephacryl S-300 of deoxycholate-treated membranes and reconstituted into liposomes by dialysis or by the Biobeads SM-2 method. Assay of the signal peptidase activity was performed with pre-human placental lactogen synthesized by the reticulocyte lysate system programmed with human placental lactogen mRNA. The signal peptidase reconstituted into liposomes showed stable activity over the temperature range of 0 to 45 degrees C; in contrast, the detergent-solubilized signal peptidase of dog pancreatic membranes was completely inactivated at the unusually low temperature of 37 degrees C. It was shown that this inactivation was due to the presence of detergent. Signal peptidase from rat liver was insensitive to a variety of protease inhibitors, like the enzyme from dog pancreas, but differed from the latter in being inhibited by chymostatin and TPCK.

Animals↗
PubMed1977

Isolation and characterization of membrane proteins responsible for attachment of polyribosomes to rough microsomal fraction of rat liver.

A protein fraction which has a high affinity for polyribosomes was isolated from rough microsomal membranes of rat liver. The mode of polyribosome binding to this fraction (R-fraction) was studied by using CsCl equilibrium centrifugation and compared with that for stripped rough microsomal membranes. The following were found. (1) The polyribosome-binding cpacity of the R-fraction was heat-labile and sensitive to trypsin, and was suppressed by increasing KCl concentration and addition of 0.1 mM-aurintricarboxylic acid. (2) Of the four subfractions obtained by gel filtration of the R-fraction on a Sephadex G-200, only the R1-fraction, eluted at the void volume, showed a high affinity for polyribosomes. The polyribosome-binding capacity of the R1-fraction decreased with time on storage at 4 degrees C. (3) The R1-fraction contained three major proteins with mol. wts. 108,000, 99,000 and 65,000.

Animals↗