1989 Fiscal Year Final Research Report Summary
Molecular biology of melatonin synthesis in pineal gland
| Project/Area Number |
63480133
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | Tokyo Metropolitan Institute for Neurosciences |
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Principal Investigator |
DEGUCHI Takeo Tokyo Metropolitan Institute for Neurosciensce, Department of Molecular, Neurobiology Laboratory Chief, 分子神経生物学研究室, 参事研究員 (20073059)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Yukiko Tokyo Metropolitan Institute for Neurosciensce, Department of Molecular, Researc, 分子神経生物学研究室, 主事研究員 (30142152)
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| Project Period (FY) |
1988 – 1989
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| Keywords | pineal gland / melatonin / N-acetyltransferase / polymorphism / point mutation |
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| Research Abstract |
Biological substances and drugs that contain amino-group are converted by N-acetyltransferase (NAT) to either bioactive substances or to their inactive form to be finally secreted from the body. Serotonin N-acetyltransferase (SNAT) that is involved in the synthesis of melatonin belongs to the first class, whereas arylamine NAT in the liver is involved in the inactivation of amine-containing drugs. In this study, we have tried to clone the cDNAs for NATs from various tissues including the pineal gland, to elucidate the relationship between structures and functions of NATs, and to detected the gene mutation associated with N-acetylation polymorphism.
1. We purified arylamine NAT from chicken liver and raised monoclonal antibody using a partially purified enzyme preparation. Homogeneous NAT protein was obtained by use of immunoaffinity chromatography and its amino acid sequences were determined.
2. Using an oligonucleotide corresponding to the amino acid sequences the cDNA clone for liver NAT was isolated and expressed in mammalian cells CHO.
3. The chicken liver cDNA was employed as a probe for the screening of the cDNA library prepared from the poly(A)-rich RNA of chicken pineal gland. Two independent cDNA clones were isolate and shown to code for arylamine N-acetyltransferases.
4. The cDNAs for NATs were isolated from the livers of rabbit and human. Two different cDNAs were detected in human liver that corresponded to the polymorphic NAT. Further analysis of the cDNAs indicated that a cDNA correlated with a high NAT activity and the other to a low NAT activity.
5. Genomic Southern blot analysis has shown that there are RFLP in the polymorphic NAT genes. Actually three different genes were detected that code for the polymorphic NAT: one gene corresponded to a high NAT activity, whereas the other two correlated with a low NAT activity.
6. Genomic Southern blot analysis has also shown that in a slow acetylator rabbit, the NAT gene was partially deleted.
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Research Products
(12 results)
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[Publications] Deguchi, T., Sakamoto, Y. Sasaki, Y., Uyemura, K.: "Arylamone N-acetyltransferase from chicken liver. I. Monoclonal antibodies, immunoaffinity purification, and amino acid sequences." J. Biological Chemistry 263, 7528-7533, 1988.
Description
「研究成果報告書概要(欧文)」より
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[Publications] Ohsaki, S., Ohtomi, M., Sakamoto, Y., Uyemura, K. and Deguchi, T.: "Arylamine N-acetyltransferase from chicken liver. II. Cloning of cDNA and expression in Chinese hamster ovary cells." J. Biological Chemistry 263, 7534-7538, 1988.
Description
「研究成果報告書概要(欧文)」より
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