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M Furuichi

Publications and source records attributed to M Furuichi.

34 records · Page 2Linked to original sources
PubMed1997

Detergent effects on the light-harvesting chlorophyll A/B-protein complex crystallization revealed by fluorescence depolarization.

Detergent effects on the pre-crystallization of light-harvesting chlorophyll a/b-protein complex (LHCII) were investigated through the fluorescence depolarization method. Stable LHCII crystals were formed in the media containing Triton X-100 (TX) or n-nonyl-beta-D-glucopyranoside (NG) and the crystallization efficients were dependent on their concentrations. The second virial coefficient of crystallizing system, estimated by the fluorescence depolarization analysis of LHCII, showed the most harmonious value under the condition yielding the most efficient crystallization of LHCII to suggest that some specific molecular interaction leading to the crystal growth would be induced according to the concentration of TX or NG.

Chlorophyll↗
PubMed1996

Requirement for two conserved cysteine residues in the Ada protein of Escherichia coli for transactivation of the ada promoter.

Cysteine residue 69 of the Escherichia coli Ada transcription factor, which accepts a methyl group from methylphosphotriester in methylated DNA, was substituted by each of 19 other amino acids. Only the mutant Ada (C69H), carrying a histidine substitution of Cys69, exhibited a limited degree of transactivating potential for the ada promoter in E. coli cells although the mutant protein was completely devoid of methylphosphotriester-DNA methyltransferase activity. Using a multicopy plasmid system for the expression of Ada protein, we have shown that Ada C69H has a transactivating capacity equivalent to that of wild-type Ada protein in the absence of an alkylating agent. This indicates that the zinc-binding capacity of histidine at residue 69 is likely to be sufficient for Ada to recognize and bind to the ada promoter. Furthermore, transactivation of the ada promoter by Ada C69H was enhanced up to 6-fold by treatment with methylating agents. An additional substitution was made with alanine in Ada C69H, replacing Cys321, the site for acceptance of a methyl group from O6-methylguanine and O4-methylthymine residues in DNA, with alanine. This renders the protein completely inactive as a methyltransferase but this derivative is constitutively active as a transactivator for the ada promoter. Therefore, acquisition of a methyl group at Cys321 apparently enhances the transactivating capacity of Ada protein on the ada promoter. We propose that the transcription-regulating function of Ada protein is under dual control by methylation of cysteine residues at positions 69 and 321; the former enhances DNA binding, while the latter enhances the transactivating capacity of the protein.

Amino Acid Sequence↗
PubMed1995

Partial purification and characterization of histidine acetyltransferase in brain of Nile tilapia (Oreochromis niloticus).

High activity of histidine acetyltransferase (HISAT) was found in the brain and the lens of Nile tilapia Oreochromis niloticus. HISAT was semi-purified 4166-fold from the brain of Nile tilapia. The affinity chromatography using a Blue Sepharose 6 FF column was very effective for purification of this enzyme. The enzyme had a broad pH optimum from pH 7.0 to pH 9.5, and did not require any divalent metal ion. The semi-purified HISAT showed a strict substrate specificity for L-histidine (and its methyl derivatives) and acetylcoenzyme A (CoASAc). The reaction velocity fits normal Michaelis-Menten kinetics with respect to both L-histidine (Km, 0.45 mM) and CoASAc (Km, 0.027 mM). Gel filtration on Superdex 200 HR indicated the molecular weight of 39,000. It was presumed that the 38.5 kDa protein, which was intensely visualized in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was a single subunit derived from HISAT.

Acetyl Coenzyme A↗
PubMed1995

Intracellular localization of 8-oxo-dGTPase in human cells, with special reference to the role of the enzyme in mitochondria.

We examined the intracellular distribution of 8-oxo-dGTPase (8-oxo-7,8-dihydrodeoxyguanosine triphosphatase) encoded by the MTH1 gene, a human mutator homologue. The activity of 8-oxo-dGTPase mainly located in cytosolic and mitochondrial soluble fractions of Jurkat cells, a human T-cell leukemia line. Electron microscopic immunocytochemistry, using a specific antibody against MTH1 protein, showed localization of MTH1 protein in the mitochondrial matrix. Activity in the mitochondria accounted for about 4% of the total activity. The specific activity in the mitochondrial soluble fraction (8093 units/mg protein) was as high as that in the cytosolic fraction (8111 unit/mg protein). The 8-oxo-dGTPase activities in cytosolic and mitochondrial soluble fractions co-eluted with MTH1 protein by anion-exchange chromatography, and the molecular mass of the mitochondrial MTH1 protein was much the same as that of the cytosolic MTH1 protein (about 18 kDa). HeLa cells expressing MTH1 cDNA showed an increased cytoplasmic signal together with a weak signal in the nucleus in in situ immunostaining of MTH1 protein, and the overexpressed MTH1 protein was recovered from both cytosolic and mitochondrial fractions. Thus, the 8-oxo-dGTPase encoded by MTH1 gene is localized in mitochondrial and cytosol.

Blotting, Western↗
PubMed1994

N-methyl-D-aspartate receptor-mediated, prolonged afterdischarges of CA1 pyramidal cells following transient cerebral ischemia in the rat hippocampus in vivo.

We previously reported the post-ischemic potentiation (PIP) of synaptic efficacy in hippocampal Schaffer collateral/CA1 responses of the rat beginning at 6-8 h following 12 min transient cerebral ischemia in vivo. The present study demonstrated that repetitive stimulation with a relatively low frequency (5 Hz, 6 s), which produced short-lasting afterdischarges (ADs; duration, 4.49 +/- 4.26 s; n = 7) in sham-controls, resulted in prolonged ADs (duration, 26.33 +/- 12.63 s; n = 6; P < 0.001) at the same period after ischemia. The PIP was not affected by 2-amino-5-phosphonovalerate (APV) administered via microdialysis at 7 h post-ischemia. The prolonged ADs in response to repetitive stimulation were, however, reversed to short-lasting ADs (duration, 7.13 +/- 1.44 s; n = 4; P < 0.02) by the same procedure, leaving the response to single stimulation unaffected. These findings suggest that, during the reperfusion period, Ca2+ influx into the CA1 pyramidal cells can be greatly increased through N-methyl-D-aspartate (NMDA) receptor-coupled ion channels if appropriately timed multiple synaptic inputs bombard these cells. Such Ca2+ influx may contribute to delayed death of CA1 pyramidal cells after transient cerebral ischemia if synaptic activity is maintained at relatively high levels during the reperfusion period.

Animals↗
PubMed1994

Genomic structure and chromosome location of the human mutT homologue gene MTH1 encoding 8-oxo-dGTPase for prevention of A:T to C:G transversion.

8-Oxo-dGTP (8-oxo-7,8-dihydrodeoxyguanosine triphosphate) is produced by active oxygen species in the nucleotide pool of the cell and can be incorporated into cellular DNA. Human cells contain enzyme activity that hydrolyzes 8-oxo-dGTP to 8-oxo-dGMP, thereby preventing occurrence of mutations, caused by misincorporation. When the cDNA for human 8-oxo-dGTPase was expressed in Escherichia coli mutT- mutant cells devoid of self 8-oxo-dGTPase activity, the elevated level of spontaneous A:T to C:G mutation frequency reverted to normal. We isolated the genomic sequence encoding the enzyme and named the gene MTH1 (for mutT human homologue). This gene is composed of at least 4 exons, spans approximately 9 kb, and is located on human chromosome 7p22.

Amino Acid Sequence↗
PubMed1993

Cloning and expression of cDNA for a human enzyme that hydrolyzes 8-oxo-dGTP, a mutagenic substrate for DNA synthesis.

8-Oxoguanine (8-oxo-7, 8-dihydroguanine) is produced in DNA, as well as in nucleotide pools of cells, by active oxygen species normally formed during cellular metabolic processes. 8-Oxoguanine nucleotide can pair with cytosine and adenine nucleotides at almost equal efficiencies, and transversion mutation ensues. Human cells contain enzyme activity, which hydrolyzes 8-oxo-dGTP to 8-oxo-dGMP, and this enzyme is responsible for preventing misincorporation of 8-oxoguanine into DNA. We purified this particular human enzyme to physical homogeneity and determined a partial amino acid sequence. We then cloned the cDNA for human 8-oxo-dGTPase and examined its nucleotide sequence. The human protein comprises 156 amino acid residues and has some sequence homology with the Escherichia coli MutT protein, which has a distinct 8-oxo-dGTPase activity. When the human cDNA was expressed in E. coli mutT- mutant cells, there was a significant amount of 8-oxo-dGTPase activity. In such cells, the frequency of spontaneous mutation was greatly reduced. We propose that the human 8-oxo-dGTPase protects genetic information from the untoward effects of endogenous oxygen radicals.

Amino Acid Sequence↗
PubMed1993

Post-ischemic potentiation of Schaffer collateral/CA1 pyramidal cell responses of the rat hippocampus in vivo: involvement of N-methyl-D-aspartate receptors.

The present study examined the functional changes in the hippocampal CA1 pyramidal cell system in vivo occurring after 12-min forebrain ischemia in the rat. A population excitatory postsynaptic potential and orthodromic population spike of CA1 pyramidal cells to stimulation of the Schaffer collaterals were potentiated at 6-8 h post-ischemia. These changes were not associated with an increase in excitability of the CA1 pyramidal cells as evaluated from the antidromic population spike induced by alveus stimulation, suggesting the presence of an increased synaptic efficacy. The post-ischemic potentiation was prevented by pretreatment with the N-methyl D-aspartate (NMDA) receptor antagonist, MK801, in a dose-dependent manner. These findings suggest that 12-min forebrain ischemia in the rat activates NMDA receptors, which results in an increase in synaptic efficacy to the CA1 pyramidal cells at 6-8 h post-ischemia.

Animals↗
PubMed1993

Impairment of hippocampal long-term potentiation following transient cerebral ischaemia in rat: effects of bifemelane, a potent inhibitor of ischaemia-induced acetylcholine release.

The present study examined the changes in the long-term potentiation (LTP) of the hippocampal Schaffer collateral/CA1 pyramidal cell system in vivo occurring after 12-min forebrain ischaemia in the rat. A population spike (PS) of CA1 pyramidal cells were recorded in 18 rats at 7 h post-ischaemia. While the threshold and amplitude of pretetanic PS were not depressed, the induction of LTP was significantly impaired following ischaemic insult (155 +/- 28% of baseline PS, mean +/- SD, n = 6, p < 0.001), as compared to sham-controls (337 +/- 84%, n = 5), without reduction of single pulse responses. Pretreatment with 4(o-benzylphenoxy)-N-methylbutylamine hydrochloride (Bifemelane; 50 mg kg-1 i.p.) slightly but significantly attenuated the impairment in LTP induction (196 +/- 23%, n = 6; p < 0.02). The findings in the present study indicate that CA1 pyramidal cells are dysfunctional in synaptic plasticity long before their death. Bifemelane has been demonstrated to inhibit acetylcholine release strongly during cerebral ischaemia but not interfere with normal cholinergic synaptic transmission. The effect of bifemelane suggests that Ach release during ischaemia is partially involved in the post-ischaemic impairment of LTP induction.

Acetylcholine↗
PubMed1992

Regulatory elements for expression of the alkA gene in response to alkylating agents.

Expression of the alkA gene in Escherichia coli is controlled by Ada protein, which binds to a specific region of the alkA promoter and enhances further binding of RNA polymerase holoenzyme to the complex. To determine the sequence recognized by the Ada protein, we introduced various base substitutions into the promoter region of alkA and examined their effects on expression of the gene, both in vivo and in vitro. Base changes within the sequence AAAGCAAA, located between positions -41 and -34 from the transcription initiation site, greatly decreased the frequencies of initiation of transcription. In footprinting experiments, the region containing this sequence was protected by the Ada protein and base changes within this sequence led to failure of binding of Ada protein to the promoter. It is likely that the Ada protein recognizes the AAAGCAAA sequence in the alkA promoter and binds to the region containing the sequence, thereby allowing ready access of RNA polymerase to the promoter. There are considerable differences between the mechanisms of action of Ada protein on the promoters of alkA and ada, even though the expression of both genes is positively regulated by Ada protein.

Alkylating Agents↗
PubMed1991

The presence and active transcription of three independent leader exons in the mouse insulin-like growth factor II gene.

The presence of multiple leader exons is one of the common features in the insulin-like growth factor II (IGFII) genes. Among them the 5' most exon sequence, rE1, was so far reported to be present only in the rat genome. We have found a rE1-homologous sequence in the mouse genome (mE1) and have isolated it by genomic cloning. The mE1 sequence was located in the 5' region of the IGFII gene and was considered to take an integral part in the mouse IGFII gene construction, just like in the rat gene. Overall homology between mE1 and rE1 regions was approx. 95%. The mE1 was actively transcribed in the newborn tissues and generated approx. 3.8 kb RNA species. Since the other two leader exon sequences were also active, producing 4.6 kb and 3.6 kb RNA species, respectively, transcription units of the mouse IGFII gene were, thus, composed of three leader exon systems.

Animals↗
PubMed1991

Growth of totally thrombosed giant aneurysm within the posterior cranial fossa. Diagnostic and therapeutic considerations.

We report a case in which growth of a totally thrombosed giant aneurysm of the posterior cranial fossa was demonstrated by computed tomography (CT) scans repeated after 4 years. A magnetic resonance (MR) image demonstrated an onion-skin-like, laminated structure within a calcified wall. The laminated structure had developed around an old thrombosis, without any communication to the flowing blood. It showed intensities indicating recent clots, revealing that the giant aneurysm had grown by recurrent intramural hemorrhage rather than by intraluminal accumulation of thrombotic materials. This case illustrates that totally thrombosed giant aneurysms still have the potential of growth.

Cerebral Angiography↗
PubMed1989

A LINE 1 sequence interrupts the rat alpha 2u globulin gene.

The rat alpha 2u globulin gene family is composed of about 25 members with closely related structure. Several of these genes are transcriptionally active in the liver of adult males. We isolated about 50 independent genomic clones from the library and determined complete nucleotide sequences of two. Although they were alike and revealed structural features prerequisite for active genes, one cloned gene was interrupted in the 6th intron with about 900 bp 5' truncated and 3'-terminal triplicated form of LINE 1 sequence flanked by 16 bp complete direct repeat originating from the 6th intron. Within the inserted LINE 1 an excellent splicing acceptor homologue with a precedent lariat junction motif and multiple polyadenylation signals was identified. Thus, the possible production of a chimeric RNA, the last exon of which would be replaced by a part of LINE 1, has to be considered.

Alpha-Globulins↗
PubMed1984

An altered DNA sequence encompassing the ras gene of Harvey murine sarcoma virus.

The DNA fragment encompassing the ras gene of Harvey murine sarcoma virus was sequenced and assigned the coding region of a transforming protein, p21, to the sequence. Examination of nucleotide sequence, taken together with the result of analysis of the ras mRNAs (1), has revealed that p21 is encoded from a continuous coding region starting with the 5' proximal initiation codon but not a processed protein. However, there were found several differences between the sequence published by Dhar et. al. (2) and ours, including 9 deletions, 7 substitutions and 2 insertions of nucleotides in the published sequence of 997 nucleotides in length. Among these, one of the substitutions occurring in the coding region resulted in amino acid replacement of glycine by alanine at position 122 of p21. The evidences are presented with some of actual gel autoradiographs.

Amino Acid Sequence↗
PubMed1979

The specific, diverse effects of purine and pyrimidine nucleoside-5'-di(tri)-3'-diphosphates on the eucaryote translation system in vitro.

All the eight 5'-di(tri)-3'-diphosphates of four common ribonucleosides were prepared by the enzymic pyrophosphoryl transfer catalysed by Streptomyces adephospholyticus ATP:nucleotide pyrophosphokinase (E.C.2.7.6.4) form dATP to the respective 5'-phosphates, and their effects on the translation of mRNAs by a wheat germ system in vitro were studied. (p) ppPupp decreased the total 14C-leucine incorporation directed by a rat liver mRNA whereas (p) ppPypp did not. With a silkworm pupa ovary mRNA, distinctly reverase results were obtained. Gel electrophoretic profiles of the translation products disclosed the mRNA species-specific stimulatory or inhibitory effects for each of the polyphosphates tested.

Adenine Nucleotides↗