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Biomedical subjects

T Harano

Publications and source records attributed to T Harano.

At least 127 records · Page 7Linked to original sources
PubMed1981

Intracellular and intramembranous localization of a protein disulfide isomerase in rat liver.

The intracellular distribution of a membrane-bound protein disulfide isomerase (PDI) in rat liver was studied by quantitative immunoprecipitation, and its microsomal localization was confirmed. The content of the enzyme was 1 to 2% of total microsomal protein, and it was almost equally distributed between rough and smooth microsomes. The enzyme was not solubilized from microsomes by high concentrations of KCl, but was readily solubilized by detergents. Since PDI in microsomes was susceptible to digestion by trypsin, at least some parts of the enzyme molecule are exposed on the outside surface of microsomal vesicles. However, the binding of antibodies to microsomal PDI and the modification of the glutamine residues of PDI molecules by transglutaminase suggested that the molecules are not extensively exposed on the surface. Solubilized PDI was unable to rebind to microsomes or to become incorporated into reconstituted membrane of detergent-solubilized microsomes, showing that the association of this enzyme with the membrane is not simply mediated by hydrophobic interaction.

Animals↗
PubMed1981

Biosynthesis and turnover of a microsomal protein disulfide isomerase in rat liver.

The biosynthesis and turnover of one type of microsomal protein disulfide isomerase (PDI) in rat liver were studied. The enzyme is predominantly synthesized by the membrane-bound ribosomes of rough endoplasmic reticulum, as judged from the results of immunological analyses of puromycin-released nascent peptides and in vitro translation products of isolated free and bound polyribosomes. Nascent peptides of PDI were released from rough microsomes by treatment with puromycin in the presence of 0.5 M KCI, which indicates that the nascent peptides of this enzyme are released on the outer surface of microsomal vesicles. This enzyme accounts for about 0.1 to 0.2% of the total protein synthesis by membrane-bound ribosomes. The half-life of PDI was about 7 days, which was significantly longer than the average half-life of microsomal proteins.

Animals↗
PubMed1981

Hemoglobin Yusa (beta 21 (B3) Asp leads to Tyr), a new abnormal hemoglobin found in Japan.

An electrophoretically slowly moving hemoglobin variant, Hb Yusa, was found in a 48-year-old man, his younger sister and mother. No clinical or hematological abnormalities were seen in the individuals heterozygous for this mutant gene. Structural studies of this abnormal hemoglobin showed a new type substitution of beta 21 (B3) Asp leads to Tyr. The percentage of the abnormal hemoglobin in the hemolysate was 40.8. The hemoglobin had neither instability nor functional abnormality.

Amino Acids↗
PubMed1980

Hemoglobin takamatsu (beta 120 (GH 3) Lys leads to Gln): a new abnormal hemoglobin detected in three unrelated families in the takamatsu area of shikoku.

A survey of hemoglobinopathies which was carried out in the Takamatsu district during the period from January to August 1979 detected six families with abnormal hemoglobins. Approximately 6010 inhabitants were screened. Three of these families had the same new Hb variant (Hb Takamatsu beta 120 Lys leads to Gln) that has not been previously reported. Existence of a blood relationship among these three families could not be established even after careful family studies. This abnormal hemoglobin was not associated with adverse symptoms and gave normal hematologic findings in the carriers. The isopropranol test was negative, oxygen affinity was within the normal range, and biosynthetic ratio in reticulocytes was around 1.0. One of the difficulties in the structural analysis of this hemoglobin was related to complete superposition of abnormal beta XT-12b,13 on a beta T-8,9 peptide in the fingerprint of the trypsin digest of aminoethylated aberrant beta X chain. This was overcome by collection of abnormal tryptic beta core (beta XT-10-13) from unmodified beta X chain, and subsequent digestion by chymotrypsin. Edman analysis of the chymotryptic peptides thus obtained successfully confirmed the substitution to be beta 120 Lys leads to Gln.

Adult↗
PubMed1978

Characterization of electron transport enzymes in the envelope of rat liver nuclei.

Nuclei and microsomes were prepared from the livers of normal, phenobarbital (PB)-treated and beta-naphthoflavone (beta-NF)-treated rats, and the contents of several enzymes in both subcellular fractions were examined. In normal rats, the enzyme activities in the nuclear fraction were about one-third of those of microsomes on a phospholipid basis. The induction of some particular enzymes by the drugs was observed with nuclei as well as with microsomes. Cytochrome P-450 and NADPH-cytochrome c reductase were increased by PB treatment and cytochrome P-448 was induced by beta-NF treatment both in nuclei and in microsomes. The extents of inhibition of nuclear enzyme activities by the antibodies against corresponding microsomal enzymes were almost the same as those of the microsomal activities. It was concluded that a microsomal type electron transport system exists in rat liver nuclei, and that nuclear drug-oxidizing activities are inducible by PB or beta-NF as their microsomal counterparts are.

Animals↗
PubMed1978

Biogenesis of endoplasmic reticulum membrane in rat liver cells. III. Biosynthesis of NADPH-cytochrome c reductase and cytochrome b5 by loosely-bound ribosomes.

Membrane-bound ribosomes were separated into two distinct classes (loosely-bound and tightly-bound ribosomes) by treatment with 0.6 M KCl, 1 mM puromycin, 0.05% DOC, or 10 mM EDTA. It was also confirmed that any one of these reagents except for EDTA dissociated the same class of ribosomes from the membrane. A population of lighter microsomal vesicles was formed from rough microsomes upon the dissociation of loosely-bound ribosomes by treatment with these chemicals. Rough microsomes were subfractionated into lighter and heavier fractions, L-rMs and H-rMs, by centrifugation using a discontinuous gradient of sucrose consisting of 1.3 M, 1.5 M, and 2.1 M solutions. It was found that L-rMs was rich in loosely-bound ribosomes, whereas H-rMs contained a high proportion of tightly-bound ribosomes. It is likely that loosely-bound and tightly-bound ribosomes are heterogeneously distributed among rough microsomal vesicles. Loosely-bound ribosomes and tightly-bound ribosomes synthesize different kinds of proteins. Two microsomal membrane proteins, NADPH-cytochrome c reductase and cytochrome b5, were exclusively synthesized by loosely-bound ribosomes, whereas serum albumin, which is a major component of the secretory proteins of hepatocytes, was synthesized only by tightly-bound ribosomes. Since the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 are released from bound ribosomes to the cytoplasmic surface of endoplasmic reticulum, while those of secretory proteins are discharged into the lumen across the membrane, the strength of the association between ribosomes and microsomal membrane seems to be correlated with the direction of release of nascent peptides.

Animals↗