Membrane-bound acid pyrophosphatase from Sulfolobus tokodaii, a thermoacidophilic archaeon: heterologous expression of the gene and characterization of the product.

Membranes of Sulfolobus tokodaii, a thermoacidophilic archaeon that grows optimally at pH 2-3, 75-80 degrees C, show the ability to hydrolyze PPi with an optimum pH of 2-3. This acid PPase is proposed to be a dolicholpyrophosphatase that participates in glycoprotein biosynthesis. In the present study, the archaeal membranes hydrolyzed isopentenylpyrophosphate and geranylpyrophosphate, compounds related to dolicholpyrophosphate, at pH 3.

Thermus kawarayensis sp. nov., a new member of the genus Thermus, isolated from Japanese hot springs.

A long-rod-shaped thermophilic microorganism, strain KW11, was isolated from a hot springs located in the Kawarayu, Gunma, Japan. Cloning and preliminary sequence analysis of 16S rDNA showed that this isolate belongs to the genus Thermus. The cells were 10-20 microm long, about 0.

Reclassification of Sulfolobus hakonensis Takayanagi et al. 1996 as Metallosphaera hakonensis comb. nov. based on phylogenetic evidence and DNA G+C content.

The taxonomic status of Sulfolobus hakonensis Takayanagi et al. 1996 was re-evaluated by fresh determinations of the 16S rDNA sequence and G+C content of the genomic DNA of the type strain, HO1-1(T). The 16S rDNA sequence of strain HO1-1(T) showed 98 % similarity to those of two Metallosphaera species and only View Article and Find Full Text PDF

[Expression of stress protein (HSP 70) in adrenalectomized mice].

We have already reported that the expression of HSP 70 in the adrenal gland was the most apparent among all organs in whole body-heated mice. In this paper, we examined the relation between the induction of HSP 70 and the adrenal gland using adrenalectomized mice. The expression of HSP 70 in some organs of adrenalectomized mice was significantly lowered in comparison with control mice.

The evolution of lipids.

Living organisms on the Earth which are divided into three major domains--Archaea, Bacteria, and Eucarya, probably came from a common ancestral cell. Because there are many thermophilic microorganisms near the root of the universal phylogenetic tree, the common ancestral cell should be considered to be a thermophilic microorganism. The existence of a cell is necessary for the living organisms; the cell membrane is the essential structural component of a cell, so its amphiphilic property is vital for the molecule of lipids for cell membranes.

Metallosphaera sedula TA-2, a calditoglycerocaldarchaeol deletion strain of a thermoacidophilic archaeon.

A spherical thermoacidophilic archaeon, strain TA-2, was obtained from acidic hot springs located in Ohwaku Valley, Hakone, Japan. This isolate is an obligate aerobic chemoorganoheterotroph that grows optimally at about 75 degrees C, pH 2.8.

Variation in molecular species of polar lipids from thermoplasma acidophilum depends on growth temperature.

Five types of molecular species of C40 isoprenoid chains, having different numbers of cyclopentane rings, were detected in the ether core lipid of Thermoplasma acidophilum. The average cyclization number of the hydrocarbon chains in the lipids increased with increasing growth temperatures.

Phylogenetic analysis of archaeal PCNA homologues.

Proliferating cell nuclear antigen (PCNA) is an essential component of the DNA replication and repair machinery in the domain Eucarya. Eukaryotes and euryarchaeotes, which belong to one subdomain of Archaea, possess a single PCNA homologue, whereas two distinct PCNA homologues have been identified from Sulfolobus solfataricus, which belongs to the other archaeal subdomain, Crenarchaeota. We have cloned and sequenced two genes of PCNA homologues from the thermoacidophilic crenarchaeon Sulfurisphaera ohwakuensis.

Characterization of caldarchaetidylglycerol analogs, dialkyl-type and trialkyl-type, from Thermoplasma acidophilum.

The structures of three kinds of phospholipids (PL-X, PL-Y, and PL-T) isolated from Thermoplasma acidophilum have been characterized. The core lipid of PL-Y was caldarchaeol, and that of PL-X was archaeol. The composition of the hydrocarbon chains of the PL-T core lipid was C20 phytane and C40 isoprenoid in a molar ratio of 2 to 1.

Sequence analysis of three family B DNA polymerases from the thermoacidophilic crenarchaeon Sulfurisphaera ohwakuensis.

Three family B DNA polymerase genes, designated B1, B2, and B3, were cloned from the thermoacidophilic crenarchaeon Sulfurisphaera ohwakuensis, and sequenced. Deduced amino acid sequences of B1 and B3 DNA polymerases have all exonuclease and polymerase motifs which include critical residues for catalytic activities. Furthermore, a YxGG/A motif, which is located between 3'-5' exonuclease and polymerization domains of family B DNA polymerases, was also found in each of the B1 and B3 sequences.

Pre-treatment with mild whole-body heating prevents gastric ulcer induced by restraint and water-immersion stress in rats.

The purpose of this study was to assess the preventive effect of pre-mild whole-body heating (WBH) on gastric ulcer induced by restraint and water-immersion stress. The ulcer index and ulcer area ratio in rats exposed to restraint and water-immersion stress were significantly decreased (p < 0.05 for both) after pre-treatment with mild WBH, compared with non-pre-treated rats.

Glucosylcaldarchaetidylglycerol, a minor phosphoglycolipid from Thermoplasma acidophilum.

A novel phosphoglycolipid (GPL-K) was isolated from Thermoplasma acidophilum (ATCC 27658). The chemical components of GPL-K were analyzed by gas liquid chromatography and GC-MS. The sugar moiety of GPL-K and its anomeric region were analyzed by NMR assignment.

Partial purification and characterization of thermostable acid phosphatase from thermoacidophilic archaeon Sulfolobus acidocaldarius.

Thermostable acid phosphatase (APase) from thermoacidophilic archaeon Sulfolobus acidocaldarius was isolated, partially purified, and characterized. The optimum pH and temperature of the enzyme for p-nitrophenylphosphate (pNPP) as a substrate were 5.0 and 70 degrees C, respectively.

Isolation and characterization of novel neutral glycolipids from Thermoplasma acidophilum.

Several novel neutral glycolipids (GL-1a, GL-1b, GL-2a, GL-2b and GL-2c) were isolated from Thermoplasma acidophilum by high-performance liquid chromatography using phenylboronic acid-silica and preparative thin-layer chromatography. The tentative structures of these lipids were characterized by the combination of gas-liquid chromatography, the methylation procedure, and (1)H-NMR and FAB-mass spectrometries. The lipophilic portion of the neutral glycolipids was composed of a simple molecular species named caldarchaeol (dibiphytanyl-diglycerol tetraether).

Isoprenoid synthesis gene for geranylgeranyl diphosphate synthase from a hyperthermophile, Pyrococcus sp. strain OT3.

Pyrococcus sp. strain OT3, a hyperthermophilic archaeon that was isolated by the authors was found to contain tetraether lipid mainly in the membrane lipid, which was quite different from the other hyperthermophiles (Masuchi et al. 1997).

Sulfurisphaera ohwakuensis gen. nov., sp. nov., a novel extremely thermophilic acidophile of the order Sulfolobales.

Three spherical thermoacidophilic archaea (strains TA-1T, TA-13, TA-14) were obtained from acidic hot springs located in Ohwaku Valley, Hakone, Japan. All the isolates are facultatively anaerobic, and grew optimally at around 85 degrees C, pH 2.0.

[Increase of cellular fibrinolysis in human lung cancer cell line by radiation: relationship between urokinase-type plasminogen activator (uPA) and metastasis and invasion].

It is well known that urokinase-type plasminogen activator (uPA) activates fibrinolysis of tumor cells and accelerates their metastasis and invasion. Human adenosquamous cell line, AOI cells, were stimulated to produce and accumulate of uPA by radiation. In AOI cells, there was relationship between uPA production and accumulation and the radiation doses.

[A new quantitative assay of heat shock protein 70 by based on an enzyme-linked immunosorbent assay method].

Heat Shock Protein 70 (HSP70) was very important stress protein. HSP70 had been detected by immunoblotting or mRNA of HSP70. Our purpose is to develop an easy quantitative assay of HSP70 and utilize clinical samples, cells, tissues etc.

[Proton-magnetic resonance spectroscopy (1H-MRS) of human plasma obtained from patients with various cancer--new detection method for common cancer].

1H-Magnetic Resonance (MR) Spectra of plasma obtained from patients with various cancer were measured. Clinical usefulness of 1H-MRS assay for detection of common cancer was examined. The following results were obtained.

[19F-MR imaging of transplanted tumor: perfluorochemicals as a fluorine tumor imaging agent].

Purpose: The purpose of our study was to detect tumor selectively using 19F-magnetic resonance imaging (MRI) and to assess Fluosol-DA, a perfluorochemical emulsion, as a tumor imaging agent for 19F-MRI.

Materials And Methods: SCC VII cells were transplanted in the right leg of mice. After 6 days, Fluosol-DA was administrated intravenously (40 ml/kg).

The structure of the core polyol of the ether lipids from Sulfolobus acidocaldarius.

The major ether-type lipid structures of Sulfolobus acidocaldarius (ATCC33909) were composed of caldarchaeol and calditoglycerocaldarchaeol. However, the characterization by nuclear magnetic resonance spectroscopy and mass spectrometry showed that the structure of calditol in calditoglycerocaldarchaeol is not nonitol, 2-(1',2',3'-trihydroxypropyl)1,2,3,4,5,6-hexahydroxyhexane, but 2-hydroxymethyl-1-(2,3-dihydroxypropoxy)2,3,4,5-cyclopentanetet raol with an ether linkage in the molecule. Such an intermolecular ether linkage was resistant to BCl3 treatment, but nonresistant to 57% HI degradation treatment conducted at 100 degrees C for 60 h, producing 2-hydroxymethyl-1,2,3,4,5-cyclopentanepentaol from calditol as reaction product.

[Thermotolerance and heat shock protein (HSP 70) in vascular endothelial cells].

Recently, it was determined that the endothelial cells of blood vessel play a very important physiological role in the regulation of blood coagulation and selective permeability. To study the thermotolerance of vascular endothelial cells, human umbilical vein endothelial cells (HUVEC) were heated at 40, 43, 45 or 50 degrees C for various lengths of time with or without preheating at 40 or 43 degrees C for 30 min. The cell viability (CV) of HUVEC decreased gradually according to heating time.