| Project/Area Number |
06670155
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
UENO Hiroshi Kyoto University, Fac.of Agriculture Associate Professor, 農学部, 助教授 (30241160)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Neurotransmitter / GABA / GAD / HDC |
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| Research Abstract |
Rat brain glutamate decarboxylase (GAD) isozyme having molecular weight of 65,000 was expressed in Saccharomyces cerevisiae having a specific activity of 50 nmol/min/mg. The expressed enzyme reacted with anti-GAD65 monoclonal antibody and showed a 65 kDa band on SDS-PAGE and Western blot analysis. Detailed culture conditions were examined.
Human histidine decarboxylase (HDC) is a close relative of GAD.In order to compare the substrate specificity of two decarboxylases, we have expressed HDC in Saccharomyces cerevisiae. The expression system exhibited 200 pmol/min/mg histamine producing activity which is far superior than any tissue samples. SDS-PAGE analysis gave 74 kDa band corresponding to precursor form of HDC.
The expression system we have developed will provide strong hand on further progress in characterizing decarboxylases.
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