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T Miyata

Publications and source records attributed to T Miyata.

At least 739 records · Page 41Linked to original sources

The nucleotide sequence of the mouse immunoglobulin epsilon gene: comparison with the human epsilon gene sequence.

We have determined the nucleotide sequence of the immunoglobulin epsilon gene cloned from newborn mouse DNA. The epsilon gene sequence allows prediction of the amino acid sequence of the constant region of the epsilon chain and comparison of it with sequences of the human epsilon and other mouse immunoglobulin genes. The epsilon gene was shown to be under the weakest selection pressure at the protein level among the immunoglobulin genes although the divergence at the synonymous position is similar. Our results suggest that the epsilon gene may be dispensable, which is in accord with the fact that IgE has only obscure roles in the immune defense system but has an undesirable role as a mediator of hypersensitivity. The sequence data suggest that the human and murine epsilon genes were derived from different ancestors duplicated a long time ago. The amino acid sequence of the epsilon chain is more homologous to those of the gamma chains than the other mouse heavy chains. Two membrane exons, separated by an 80-base intron, were identified 1.7 kb 3' to the CH4 domain of the epsilon gene and shown to conserve a hydrophobic portion similar to those of other heavy chain genes. RNA blot hybridization showed that the epsilon membrane exons are transcribed into two species of mRNA in an IgE hybridoma.

Amino Acid Sequence↗

Evolutionary changes of nucleotide sequences of papova viruses BKV and SV40: they are possibly hybrids.

Complete nucleotide sequences were compared between papova viruses BKV and SV40 and the degrees of sequence divergences were compared between structurally and/or functionally different segments or genes in details. It was shown that the rate of synonymous substitution is not only very high but also approximately uniform among different genes in these viruses as in eukaryotic genes examined to date. While all the non-coding regions including the intron showed marked sequence preservation which is in sharp contrasted with the case of eukaryotic genes where the large bulk of non-coding regions evolve at a rate as rapidly as that of synonymous substitution. It is remarkable that a long continuous stretch of sequence including the putative VPX gene and a 5' half of VP2 gene showed strong homology between BKV and SV40. A close examination of the pattern of base substitutions revealed that this unusual homology was derived by recombination between the two viruses during their evolution. On the basis of the pattern of base substitutions and the bias in code word utilization, we also showed that the putative VPX gene actually could code for a functional polypeptide. In papova viruses, the 3' terminal sequence of VP2/3 gene overlaps with the 5' terminal sequence of VPI gene. The pattern of base substitutions in the overlapping segment was examined in detail in comparison with those in the non-overlapping portions of VP2/3 and VP1 genes. It was shown that the evolutionary mode of the overlapping genes is in good agreement with our previous prediction.

BK Virus↗

Plasminogen Tochigi: inactive plasmin resulting from replacement of alanine-600 by threonine in the active site.

Structural studies on a hereditarily abnormal plasminogen, plasminogen Tochigi, have been performed to identify the difference responsible for its lack of proteolytic activity. The plasminogen sample used was from a heterozygote and thus consisted of apparently equal amounts of normal and defective plasminogen molecules. Amino acid sequence analysis of a tryptic peptide isolated from the abnormal plasminogen indicated that Ala-600 (equivalent to Ala-55 in the chymotrypsin numbering system) had been replaced by Thr. No other substitutions in the active-site residues--namely, His-57, Asp-102, and Ser-195--were found. Molecular models for chymotrypsin and the bovine trypsin-pancreatic trypsin inhibitor complex indicate that Ala-55 is very near the active-site His. The Thr at position 55 in plasminogen (plasmin) Tochigi may perturb His-57 such that the proton transfers associated with the normal catalytic process cannot occur in the abnormal plasmin.

Alanine↗

Nucleotide sequences of gene segments encoding membrane domains of immunoglobulin gamma chains.

The nucleotide sequences of the exons encoding membrane-bound IgG1, IgG2a, and IgG2b of mouse were determined and compared with the sequence of membrane-bound IgM. The sequences indicate that membrane-bound gamma chains bear an additional 71 residues at the COOH termini, including 17-residue acidic, 26-residue hydrophobic, and 28 residue hydrophilic portions. The hydrophobic portion, which seems to be anchored in the lipid bilayer of the membrane, is highly conserved among membrane-bound mu and gamma chains. We propose the presence of a membrane protein that recognizes the conserved hydrophobic segment and anchors the membrane-bound immunoglobulin. Comparison of the nucleotide sequences revealed another example of intervening sequence-mediated domain exchange around one of the membrane exons.

Amino Acid Sequence↗

Nucleotide sequence of the structural gene for colicin E1 and predicted structure of the protein.

We have determined the nucleotide sequence of the structural gene for colicin E1, which consists of 1,566 base pairs. The amino acid sequence (522 residues) of the protein was derived from the DNA sequence, and the molecular weight was calculated to be 57,279. From the analysis of the predicted secondary structure, there appear to be three consecutive long alpha-helices in the NH2-terminal half of the polypeptide, spanning 40, 100, and 35 amino acid residues. In addition, there is a polypeptide region near the COOH terminus that shows homology to the NH2-terminal signal portions of outer membrane lipoprotein in Escherichia coli and beta-lactamase in Bacillus licheniformis. Most of the homologous amino acids are located in the region where either alpha-helix or beta-sheet would be expected to occur, as determined from the amino acid sequence. These characteristics of the predicted protein structure might correspond to properties of colicin E1 as an ionophore in its antimicrobial action and also as an exported protein during its induced synthesis.

Amino Acid Sequence↗

Protective effect of immediate inoculation of a live varicella vaccine in household contacts in relation to the viral dose and interval between exposure and vaccination.

The protective effect on 45 household contacts of immediate inoculation of a live varicella vaccine was examined in relation to the viral dose and days after exposure to varicella. The viral doses used were 300 PFU to 15,000 PFU, and the intervals between exposure and vaccination were 0 to 5 days. All 30 contacts given viral doses of 800 to 15,000 PFU within 3 days after exposure were protected from clinical varicella. With lower viral doses of 300 to 600 PFU within 3 days after exposure, 6 to 10 contacts developed rashes 13 to 25 days after vaccination, but all showed mild symptoms. When vaccine was given 5 days after exposure, even with a dose of 1,500 to 4,000 PFU, typical varicella symptoms appeared 6 to 9 days after vaccination. These results suggest that vaccination of household contacts with viral doses of over 500-800 PFU within 3 days after exposure is effective in preventing manifestation of clinical varicella and that clinical symptoms may be modified to a mild form by vaccination with even a lower viral dose when given within 3 days after exposure.

Chickenpox↗

A new method for evaluating mucolytic expectorant activity and its application. II. Application to two proteolytic enzymes, serratiopeptidase and seaprose.

Using our new method described in a preceding paper, in vivo effects of two proteolytic enzymes such as serratiopeptidase (SER) and seaprose (SAP) on sputa collected from bronchitis rabbits were examined. SER (20 mg/kg) and SAP (30 mg/kg) significantly reduced the viscosity of sputum (P less than 0.05) at the 1-3-h periods and the 4-6-h periods, respectively, after intraduodenal administration. 50 mg/kg of SER also significantly decreased not only viscosity (P less than 0.001) but also amount of freeze-dried substance (P less than 0.05) of sputum at the 1-3-h periods, but SAP did not affect the amount of dried substance. Both enzymes significantly increased the volume of sputum, probably as the result of liquefaction. Thus, mucolytic expectorant activity of both enzymes can be demonstrated first by the reduction in viscosity and next of the increase in volume of sputa. However, the decrease in amount of freeze-dried substance is not always in accord with the reduction viscosity.

Animals↗

A new method for evaluating mucolytic expectorant activity and its application. I. Methodology.

A new method for evaluating in vivo effect of mucolytic expectorants was devised. In this report, however, the focus is placed on methodology but not on pharmacology. Rabbits with subacute bronchitis were prepared by long-term exposure to minute amounts of SO2 gas. The sputum was quantitatively collected from the animal through a tracheal cannula at suitable time intervals according to the Perry and Boyd's method. As a preliminary study, changes in volume, viscosity and amounts of freeze-dried substance of sputa were examined after intraduodenal administration of water instead of mucolytic expectorants, the effects of which will be described in the subsequent paper, but no significant changes occurred with this test.

Animals↗

Effects of anesthetics on piperidine levels in mouse brain.

Piperidine is one of the biogenic amines possessing potent pharmacological activity. Recent interest has focused on its possible role as an endogenous hypnogenic substance. Using a mass fragmentographic technique with deuterium-labelled piperidine as an internal standard, piperidine concentrations in brains of waking and deeply anesthetized mice were analyzed to compare piperidine levels in the brain under distinctly different states of consciousness. A rapid and significant increase in piperidine concentrations was found in the brain but not in blood of mice anesthetized with any one of pentobarbital, urethane, ether and halothane. The results, showing that CNS depression is accompanied by accumulation of piperidine in the brain, are consistent with the idea that piperidine may have a close connection with the mechanisms controlling the level of consciousness.

Anesthetics↗

The complete nucleotide sequence of mouse immunoglobin gamma 2a gene and evolution of heavy chain genes: further evidence for intervening sequence-mediated domain transfer.

We have determined the complete nucleotide sequence (1990 base pairs) of mouse immunoglobulin gamma 2a gene, and compared it with the sequences of other gamma subclass genes so far sequenced, i.e. gamma 1 and gamma 2b genes. Divergence of the nucleotide sequence between a compared pair of the gamma genes varies extensively among different segments of the gene. For example, comparison of the gamma 2a and gamma 2b genes has revealed a remarkable homology in a long continuous segment (about 900 bases) that covers from the 3' portion of the first intervening sequence to the third intervening sequence. However, there is no particular segment of the gamma gene that is conserved universally among the three gamma genes. These findings suggest that, during their evolution, segments of the gamma genes had been scrambled between different subclass genes through recombinations within intervening sequences, thus providing further evidence for the intervening sequence-mediated domain transfer hypothesis. We have discussed several possible phylogenic trees which can explain the difference of divergence in various segments of the gamma genes.

Animals↗