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Biomedical subjects

K Mitsui

Publications and source records attributed to K Mitsui.

At least 217 records · Page 12Linked to original sources
PubMed1981

Purification and properties of fMet-tRNAf deacylase from Escherichia coli.

A formylmethionyl-tRNAf deacylase has been purified about 330-fold from a crude initiation factor preparation (1 M NH4Cl ribosomal wash) from Escherichia coli Q13. The enzyme was nearly homogeneous and had an apparent molecular weight of 24 000. Rat liver methionyl-tRNAf and E. coli methionyl-tRNAm were not hydrolyzed significantly by the enzyme under standard conditions. Q beta RNA- and AUG(A)n-directed polypeptide synthesis was inhibited by the enzyme. The inhibition was at the level of initiation of polypeptide synthesis. The enzymatic activity was inhibited by various factors necessary for polypeptide synthesis. The activity was inhibited more by NH4Cl and spermidine than by Mg2+, GTP and ATP. The complex of formylmethionyl-tRNAf, initiation factor 2 and GTP was resistant to enzymatic hydrolysis, and the resistance was enhanced by the addition of AUG and ribosomes to the above reaction mixture.

Acyltransferases↗
PubMed1981

Removal of beta subunit of the eukaryotic polypeptide chain initiation factor 2 by limited proteolysis.

It is generally considered that the eukaryotic polypeptide chain initiation factor 2 (eIF-2) from rabbit reticulocytes consists of three nonidentical subunits termed alpha, beta, and gamma, in order of increasing molecular weight. However, a recent report [Stringer, E. A., Chaudhuri, A., Valenzuela, D. & Maitra, U. (1980) Proc. Natl. Acad. Sci. USA 77, 3356-3359] suggested that this factor is made up of only two subunits. In this paper we show that limited proteolysis of rabbit reticulocyte eIF-2 leads to loss of the beta subunit. This modified eIF-2 has the same activity as the native factor in promoting ternary (eIF-2.GTP.Met-tRNAi) and 40S (eIF-2.GTP.Met-tRNAi.40S ribosome) initiation complex formation. Like native eIF-2, the protease-treated factor can restore translation in heme-deficient lysates. On the other hand, the treated factor is less stable than the native protein.

Animals↗
PubMed1981

The mechanism of preferential synthesis of poly[r(purine)] in the transcription of poly[d(purine)] . poly[d(pyrimidine)] by T. thermophilus RNA polymerase.

The mechanism of preferential transcription on poly[d(purine)] . poly[d(pyrimidine)] was investigated using RNA polymerase of T. thermophilus HB8. Though the machinery for initiation is lacking, the core enzyme has the latent ability to synthesize poly[r(pyrimidine)] as well as poly[r(purine)]. The holoenzyme can synthesize poly[r(purine)] in the usual manner. Poly[r(pyrimidine)] synthesis by the holoenzyme is, however, forbidden. These results suggest that the sigma factor plays a crucial role in this preferential transcription, and that this preferential transcription may be useful as a model for the sense strand recognition. Various results led us to the hypothesis that the high affinity site for the poly[d(pyrimidine)] strands on the enzyme plays a very important role.

Adenosine Triphosphate↗
PubMed1981

The structure-hemolysis relationship of oleanolic acid derivatives and inhibition of the saponin-induced hemolysis with sapogenins.

Chikusetsusaponin IV and V, whose genin is oleanolic acid, exhibited weak hemolytic activities. Removal of glucose residue at position 29 of chikusetsusaponin V by partial hydrolysis increased the activity more than 30-fold. Methylation of the carboxyl group at position 28 increased the activity furthermore by about 10-fold, showing HD50 value of 3.77 microM. On the other hand, removal of the sugar chain at position 3 of chickusetsusaponin V by partial hydrolysis completely lost the activity. These facts suggest that the sugar chain at position 3 of oleanolic acid is essential but that at position 29 is pernicious for the activity. The cytolytic agents, whose target has been regarded as membrane cholesterol, were inactivated not only by cholesterol but also by sapogenins such as oleanolic acid, gitogenin and hederagenin. Among saponins tested, akebia saponin B and C were inactivated by cholesterol, but not by the genins, probably because their affinities for the genins are too low to form complexes.

Anti-Bacterial Agents↗
PubMed1980

Purification and some properties of tetanolysin.

Tetanolysin was purified from the culture fluid of a strain of Clostridium tetani by ammonium sulfate fractionation, acetone precipitation and repeated gel filtration. Two hemolysins with different molecular weights were separated by gel filtration, and the smaller one, tetanolysin, was further purified. The purification raised the specific activity of tetanolysin 1,050-fold to 500 HU/micrograms of protein. The purified preparation gave a single, relatively broad band on polyacrylamide gel electrophoresis, in which the activity was roughly parallel with the protein concentration. However, on sodium dodecylsulfate-gel electrophoresis it gave two bands with nearly equal amounts of proteins, showing molecular weights of 53,000 and 48,000 +/- 3,000. Furthermore, isoelectric focusing revealed four peaks of the activity whose isoelectric pHs were 6.1, 5.6, 5.3, and 6.6 in decreasing order of the activity. These findings suggest that the preparation contains four hemolysins with different pIs, which are classifiable into two groups by molecular size. The preparation was completely free of tetanus neurotoxin and proteases. Tetanolysin was more strongly inhibited by cholesterol and more rapidly absorbed onto erythrocytes than theta-toxin of Cl. perfringens.

Cholesterol↗