| Project/Area Number |
13670904
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Dermatology
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| Research Institution | Kinki University |
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Principal Investigator |
KAWADA Aklira Kinki University, School of Medicine, Associate Professor, 医学部, 助教授 (90160986)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHARA Hidenari Ibaraki University, School of Medicine, Professor, 農学部, 教授 (30122063)
TEZUKA Tadashi Kinki University, School of Medicine, Professor, 医学部, 教授 (20013964)
ARAKANE Yoshinori Kinki University, School of Medicine, Lecturer, 医学部, 講師 (40232045)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Peptidylarginine deiminase / human hair cell / PAD T4 / cDNA / expression plasmid / trichohyaline / monoclonal antibody / immunohistochemistry / ペプチジルアルギニンデイミナーゼ / ヒト毛細管 / 大腸菌発現 / Two-cistran / ヒト毛嚢 / cDNAクローニング |
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| Research Abstract |
The hair is composed of inner and root sheath, cuticle, and medulla cells which are differentiated from hair mother cell. The terminal differentiation of hair mother cell is common in tracing epidermal basal cells, but the biochemical study regarding the process of hair differentiation is very few.
Peptidylarginine deiminase(PAD) is a post-translational modification enzyme which gives the electrostatic change by converting basic region of the target protein into the radio valence, causing the different interaction between target proteins and another protein. Since the first finding of G.E Rogers in 1977, that enzymatic activity converts arginine residue of the protein to the citrulline residue in extract of guinea pig hair follicle, biochemical studies of PADs were advanced in the various tissues. However the research on the PAD of the hair follicle is seldom carried out. In this study, we prepared the human hair cells by the primary culture method, and the RNA, and then cloned the enti
… More
re cDNA of the human hairy cell PAD(PAD T4) by homologue polymerase chain reaction. The complete amino acid sequence was clarified by the nucleotide sequence analysis. We constructed the human PAD T4 expression plasmid in an Echerichia coli and purification and enzymatic properties of the recombinant enzyme were clarified. Especially, the role of which the hairy cell peculiar protein(trichohyalin) is important for the regulation of the hair construct and the citrulline residues were found in the trichohyalin, that the trichohyalin is a candidate target for PAD T4. In this study we proved that PAD T4 modified the trichohyalin effectively. Next, we made a monoclonal antibody against specific for PAD T4 is and we examined the cellular distribution of PAD T4 in the hair follicle. The immunohistochemisty showed that the PAD T4 specifically localized in the inner root sheath in the hairy cells. The knowledge which proved that the above-mentioned trichohyalin also exists in this organization locally and that this enzyme has made the trichohyalin to be a target. Less
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