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Biomedical subjects

T Tezuka

Publications and source records attributed to T Tezuka.

At least 145 records · Page 8Linked to original sources
PubMed1989

[The formation of monoclonal antibody to the cell membrane region of the stratum corneum and keratohyaline granules of newborn rat].

We have developed a monoclonal antibody to the cell membrane region of the stratum corneum and keratohyalin granules using rat hematoxylin stainable proteins as an antigen. These hematoxylin stainable proteins were partially purified utilizing preparative isoelectric focusing from 50 mM Tris-HCl buffer soluble fraction of newborn rat epidermis. The developed monoclonal antibody located in both sites of the cell membrane region of the stratum corneum and keratohyalin granules (Ted-R-3). In an immunoblotting study using two-dimensional electrophoresis, a single protein which had a molecular weight of 55 kd reacted. The amino acid analysis of the affinity purified antigen showed that glycine, serine and glutamic acid residues comprised 20.4%, 18% and 12.6%, respectively. However, histidine and cystine residues comprised only 2.8% and 0.12%, respectively.

Amino Acids↗
PubMed1987

Similarity and dissimilarity in the stereogeometry of the active sites of thrombin, trypsin, plasmin and glandular kallikrein.

The relationship between chemical modifications of arginine derivatives and inhibitory activity to trypsin, plasmin and glandular kallikrein was investigated comparing with that of thrombin and concluded as follows: The hydrophobic binding pocket, which has been reported previously to be stereogeometrically very similar in trypsin and thrombin, corresponded to the length of ethylpiperidine. Concerning the site (termed the P site) next to the hydrophobic binding pocket, there were large differences in stereogeometry between trypsin and thrombin; the binding site of trypsin extended further to allow propyl and phenyl group attached to piperidine, while that of thrombin would be much narrower and unable to allow them. The P sites of plasmin and glandular kallikrein resembled that of trypsin in being able to allow phenyl group. To substantialize the hydrophobic binding pocket and the P site, a (2R, 4R)-MQPA-trypsin complex model was generated using the results of X-ray crystallography of (2R, 4R)-MQPA and BPTI-trypsin complex by calculation to minimize van der Waals contacts, and it was of great use for understanding the geometry of the active sites of trypsin, thrombin, plasmin and glandular kallikrein.

Animals↗
PubMed1987

The cystine-rich envelope protein from human epidermal stratum corneum cells.

A cystine-rich protein has been extracted from the membrane region of stratum corneum cells with 50 mM Tris-HCl buffer, pH 7.3. The protein has been purified by molecular sieve chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and preparative isoelectric focusing. An antibody, raised to the purified protein, was located in the cell membrane region in the stratum spinosum, stratum granulosum, and the inner part of the stratum corneum. In addition, the antibody also reacted with the protein extracted from the outer part of the stratum corneum by the immunoblotting method. Amino acid analysis of the protein revealed a distribution of 4.3% cystine residues, 9% lysine, 18.5% glycine, and 12.6% glutamic acid residues. The isoelectric point was found to be 4.8 and the molecular weight of the protein was 16,000.

Antibodies↗
PubMed1987

Human hematoxylin-stainable protein of keratohyalin granules origin. I. Extraction and purification.

A human hematoxylin-stainable protein (HSP) was extracted from the massive stratum corneum of epidermal cysts. This protein was purified in two steps; first, through preparative isoelectric focusing, and, second, by affinity column chromatography bound with the specific, monoclonal antibody to keratohyalin granules (Ted-H-1). In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified HSP consisted of two proteins (70 and 62 kilodaltons [kd]), but only the 62-kd protein was detected in the 15,000 g supernatant fraction of the extract using the immunoblotting technique. The amino acid composition of the purified HSP included 30% glycine, 15% serine, 12% glutamic acid, and 4% ornithine, but only 2.3% histidine. Using the indirect immunofluorescent technique, observation showed that the monoclonal antibody, Ted-H-1, to the HSP, formed as a result of the partially purified antigen, was located in three places: 1) the keratohyalin granules; 2) in the cell membrane region of the lower part of the stratum corneum of the skin samples (forearm, cheek, and back); and 3) the keratohyalin granules in the follicular epithelium and on the trichohyalin granules. Reaction product was not seen in either the acrosyringium or in the plantar epidermis. As two positively reacted proteins with the Ted-H-1 were detected in the Tris-HCl extract from the plantar stratum corneum by the immunoblotting assay, however, the above negative result in the indirect immunofluorescent studies may be due to the masking of the antigenic site of keratohyalin granules in vivo. This hematoxylin-stainable protein was synthesized as one component of the keratohyalin granules in the stratum granulosum and may have been transferred to the stratum corneum cell membrane region.

Amino Acids↗