Hisaya Iida
Gifu University
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Biochimica et Biophysica Acta | 1974
Yoshinori Nozawa; Hisaya Iida; Hirobumi Fukushima; Kazuo Ohki; Shun-ichi Ohnishi
The fatty acid distribution pattern of lipids extracted from different subcellular components of Tetrahymena pyriformis was found to be significantly different from one type of membrane to another. The growth-temperature shift caused alterations in fatty acid composition. The ratio of palmitoleic to palmitic acid, especially, showed a sharp linear decline with increase of temperature in all of the membrane fractions. The spin labels were rapidly incorporated into Tetrahymena membranes. The order parameter of 5-nitroxide stearate spin label incorporated into various membrane fractions was found to be different for the different membrane fractions, suggesting the following order of the fluidity; microsomes > pellicles > cilia. The fluidity of the surface membranes, cilia and pellicles isolated from Tetrahymena cells grown at 15°C was noticeably higher than that of the membranes from cells grown at 34°C but was not so different with microsomal fractions. The motion of the spin label in the pellicular membrane was more restricted than in its extracted lipids, thus indicating the assumption that in Tetrahymena membranes the proteins influence the fluidity. It was also suggested that a sterol-like triterpenoid compound, tetrahymanol, which is principally localized in the surface membranes, would be involved in the membrane fluidity.
Biochimica et Biophysica Acta | 1975
Yoshinori Nozawa; Hirobumi Fukushima; Hisaya Iida
Tetrahymena pyriformis WH-14 cells were grown in the medium supplemented with ergosterol (1 mg/100 ml) and the effects of replacement of tetrahymanol by ergosterol upon the lipid composition in the surface membranes (cilia and pellicles) were examined. 1. By scanning and freeze-etch electron microscopy it was suggested that exogenous ergosterol would be inserted into the lipid regions in the surface membranes. Although freeze-etched faces of filipin-treated membranes containing the native tetrahymanol showed a random distribution of 85-a protein particles, the ergosterol-replaced membranes after the same polyene treatment revealed the marked ultrastructural alterations on the fracture faces. 2. The replacement of tetrahymanol in membranes by ergosterol induced a profound alteration in the phospholipid class composition and a marked increase in phosphatidylethanolamine with a compensatory decrease in phosphatidylcholine and 2-aminoethylphosphonolipid. 3. There are significant and quantitative but not qualitative changes in the fatty acid composition of total lipids from the ergosterol-replaced membranes. There are also increases in saturated and decreases in unsaturated fatty acids. Phosphatidylethanolamine acyl chains particularly become more saturated, as compared with two other phospholipids, in ergosterol-replaced pellicles. This increase in saturation is due to an appreciable increase in C14:0, C16:0 and iso-C17:0, and a decrease in C18:1(delta9), C18:2(delta9,12) and C18:3(delta6,9,12). 4. These results suggest that profound alterations in phospholipids as well as in their fatty acyl chains are required to modify the overall membrane lipid composition for the maintenance of proper membrane fluidity. Our data would also support the thesis that polat head groups are involved in the membrane lipid organization and that sterols interact selectively with phospholipid molecules containing the appropriate fatty acyl chain composition in biological membranes.
Clinica Chimica Acta | 1974
Yoshinori Nozawa; T. Noguchi; Hisaya Iida; H. Fukushima; Takashi Sekiya; Yasufumi Ito
Abstract Erythrocyte membrane proteins of a hereditary spherocytosis patient with a severe anemia were analysed by SDS-disc electrophoresis according to the method of Fairbanks et al. A defective protein band IVb was found. Surface features of the patients red cells were examined by scanning electron microscopy which showed many deformed erythrocytes with peculiar shape. The lipid composition of erythrocyte membranes from hereditary spherocytosis was not changed.
Biochimica et Biophysica Acta | 1973
Yoshinori Nozawa; Hirofumi Fukushima; Hisaya Iida
Abstract A procedure was developed for isolation of macronuclei and nuclear membranes from the ciliated protozoan Tetrahymena pyriformis E, and the lipid composition of the isolated nuclear membranes was determined. This method involves cell lysis with octanol, separation of the nuclear membrane with 0.2 M phosphate–1M NaCl and purification on a discontinuous sucrose gradient. By phase-contrast and electron microscopic examinaton, our preparations were pure and preserved the typical nuclear membrane morphology: inner and outer nuclear membranes, and nuclear pore complexes. As for lipid distribution, the three major phospholipids in the membranes were phosphatidylcholine (31.0%), phosphatidylethanolamine (26.1%) and 2-aminoethylphosphonolipids (23.3%) and the molar ratio of a sterol-like lipid, tetrahymanol to phospholipid phosphorus was 0.036. These results were compared to other membrane fractions of Tetrahymena.
Biochimica et Biophysica Acta | 1978
Hisaya Iida; Toyozo Maeda; Kazuo Ohki; Yoshinori Nozawa; Shun-ichi Ohnishi
Abstract Transfer of phosphatidylcholine molecules between different membrane fractions of Tetrahymena pyriformis cells grown at 15, 27 and 39.5°C was studied by electron spin resonance (ESR). Microsomes were labeled densely with a phosphatidylcholine spin label and the spin-labeled microsomes were incubated with non-labeled cilia, pellicles or microsomes. The transfer of the phosphatidylcholine spin labels was measured by decrease in the exchange broadening of the electron spin resonance spectrum. In one experiment, the lipid transfer was measured between 32P-labeled microsomes and non-labeled pellicles by use of their radioactivity. The result was in good agreement with that by ESR. The fluidity of the membrane was estimated using a fatty-acid spin label incorporated into the membranes. Transfer between lipid vesicles was also studied. The results obtained were as follows: (1) The transfer between sonicated vesicles of egg- or dipalmitoyl phosphatidylcholine occurred rapidly in the liquid crystalline phase, with an activation energy of 20 kcal/mol, whereas it hardly occurred in the solid crystalline phase. (2) The transfer rate between microsomal membranes increased with temperature, and an activation energy of the reaction was 17.8 kcal/mol. (3) The transfer from the spin-labeled microsomes to subcellular membranes of the cells grown at 15°C was larger than that to the membranes of the cells grown at 39.5°C. The membrane fluidity was larger for the cells grown at lower temperature. (4) Similar tendency was observed for the transfer between microsomal lipid vesicles prepared from the cells grown at 15°C and at 39.5°C. (5) The transfer from microsomes to various membrane fractions increased in the order, cilia
Biochimica et Biophysica Acta | 1984
Yoshihito Yawata; Kosuke Miyashima; Takashi Sugihara; Naoki Murayama; Saichi Hosoda; Shigeru Nakashima; Hisaya Iida; Yoshinori Nozawa
In a patient with lecithin: cholesterol acyltransferase deficiency, free cholesterol was markedly increased, and esterified cholesterol was diminished. In the patients plasma, an increase in phosphatidylcholine (PC) and a decrease in sphingomyelin were observed. Concomitantly, an increase in a shorter acyl chain 16:0 was noted in PC, sphingomyelin and phosphatidylethanolamine (PE). In contrast to these results, longer chains such as 22:0 and 24:0 were decreased, especially in sphingomyelin. Unsaturated double bonds such as 18:1 was also increased in PC and PE. In the red-cell membrane lipids, the increase in free cholesterol was counteracted by an increase in PC and by a decrease in sphingomyelin and PE, reflecting changes in the patients plasma lipids. Increased 16:0 (in PC) and decreased 18:0 and 24:0 were observed. The increased plasma free cholesterol due to metabolic defect (lecithin: cholesterol acyltransferase deficiency) led to decreased red-cell membrane fluidity. This effect appeared to be counteracted by changing phospholipid composition (increased PC and decreased sphingomyelin and PE), by increasing shorter chains (16:0), by decreasing longer chains (18:0 and 24:0) and by increasing unsaturated double bonds (18:2). These results can be interpreted as a self-adaptive modification of lecithin: cholesterol acyltransferase deficiency-induced red-cell membrane abnormalities, to maintain normal membrane fluidity. This speculation was supported by the ESR spin-label studies on the patients membrane lipids. The normal order parameters in intact red cells and in total lipid liposomes were decreased if cholesterol-depleted membrane liposomes were prepared. Thus, the hardening effect of cholesterol appeared to be counteracted by the softening effects described above. Overall membrane fluidity in intact red cells of the lecithin: cholesterol acyltransferase-deficient patient was maintained normally, judged by order parameters in ESR spin-label studies.
Biochemical Medicine | 1984
Hisaya Iida; Yoshiki Takashima; Satoshi Maeda; Takashi Sekiya; Masaka Kawade; Masahiko Kawamura; Yukio Okano; Yoshinori Nozawa
Scanning electron microscopic observation revealed that there were wide variations including typical acanthocytes in morphology of erythrocytes from a patient with abetalipoproteinemia. The erythrocyte membrane phospholipids and cholesterol contents from a patient was higher by 25% compared to an age-matched control subject. Analysis of phospholipid composition of red blood cells showed an increase of sphingomyelin (25.1----30.1%) with a concomitant decrease of lecithin (27.5----21.0%). Thus, the sphingomyelin/lecithin ratio was increased dramatically (0.91----1.43). As for fatty acyl chain composition of main phospholipids, an increased percentage of palmitic acid and docosahexaenoic acid and a decreased proportion of arachidonic acid and lignoceric acid were observed for sphingomyelin. There was an increment of palmitic acid which was accompanied with a decrease of linoleic acid in lecithin. On the other hand, no significant difference was shown in the fatty acid composition of phosphatidylethanolamine and phosphatidylserine plus phosphatidylinositol between a patient and control.
Biochemical Medicine | 1982
Hisaya Iida; Atsushi Imai; Yoshinori Nozawa; Tokuji Kimura
Abstract The mechanism of lipid peroxidation in biological membranes has been intensively investigated with liver microsomes (1). On the other hand, much attention has been focused on lipid peroxidation from physiological and pathological aspects, including aging (2), diabetes melitus (3), stroke (4), and liver disease (5). It is believed that lipid peroxidation results in membrane damage (6–8). Wills (9) observed a loss of glucose-6-phosphatase activity during lipid peroxidation. We found that the degradation of cytochrome P -450 in bovine adrenocortical mitochondria parallels the increase in lipid peroxidation activity (10). In this communication, we wish to report to report that arachidonic acid is the best substrate for lipid peroxidation reaction among unsaturated fatty acids in bovine adrenocortical mitochondria.
Endocrine Research | 1979
Hisaya Iida; Tokuji Kimura
Lipid peroxidation of adrenocortical mitochondria and microsomes was greatly stimulated by addition of 1.0 mM or less ferric ions. In the presence of NADPH-yielding system, the formation of corticosterone from endogeneous cholesterol and exogeneous deoxycorticosterone was inhibited as the concentrations of iron increased. Of interest is the fact that 0.5 mM ferric ion-mediated lipid peroxidation was completely abroagated upon addition of 2 mM calcium ions. Accordingly, protected from the peroxidative damage.
Clinica Chimica Acta | 1979
Yukio Okano; Hisaya Iida; Taeko Yamauchi; Takashi Sekiya; Hideaki Kuwabara; Goto Masako; Yoshinori Nozawa
Biochemical and ultrastructural studies of red blood cell membranes from seven patients with congenital biliary atresia have been performed. Scanning electron microscopic observations revealed that more than half of these cells were of abnormal shapes, such as target, spur and cup-formed cells. By freeze-fracture electron microscopy, membrane particle-free smooth areas were noted in the fracture faces. In addition, the number of membrane-associated particles was 20% less than those in control subjects. The lipid analysis of red blood cells showed a significant increase in phospholipid and a marked increase in cholesterol content. The increase of phospholipid content was primarily caused by the increase of lecithin. The acyl chain analysis of erythrocyte lecithin demonstrated an increase in palmitic, palmitoleic and oleic acid, and a decrease in stearic and linoleic acid. These observations are similar to those of acquired biliary obstruction. The fluidity of erythrocyte membrane lipid, studied by a fluorescence technique using fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH), was found to be less in an individual with congenital biliary atresia than in the control subject.