1995 Fiscal Year Final Research Report Summary
The study of signal transduction mechanism of angiotensin II receptor by mean of site-directed mutagenesis
| Project/Area Number |
06660107
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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 |
応用微生物学・応用生物化学
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| Research Institution | Tottori University |
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Principal Investigator |
YAMANO Yoshiaki Tottori University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (00182593)
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| Co-Investigator(Kenkyū-buntansha) |
MORISHIMA Isao Tottori University, Faculty of Agriculture, Professor, 農学部, 教授 (30032296)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Angiotensin II / Receptor / Site-directed mutagenesis / cDNA |
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| Research Abstract |
Ligand binding of rat angiotensin II receptor (AT1A)
The molecular interaction involved in the ligand binding of rat angiotensin II receptor (AT1A) was studied by site-directed mutagenesis and receptor model building. Five conserved residues in AT1A,R167, K199, W253, F259 and D263 were substituted with alanine, individually. These amino acid residues appeared to provide binding sites for angiotensin II.Binding of a nonpeptide angiotensin II antagonist was also reduced for R167 mutant. R167 is concerned for receptor conformation.
G-protein coupling of rat angiotensin II receptor (AT1A)
G-protein coupling in carboxyl-terminal tail of rat angiotensin II receptor was studied by site-directed mutagenesis. Deletion mutants, which have stop codons at K310 (K310DELTA) or at K318 (K318DELTA), have angiotensin II binding affinity similar to wild type receptor. In case of K318DELTA, both the effects of GTPgammaS and the IP3 production was retained in response to angiotensin II,but K310DELTA had neither of the effects. It is concluded that G-protein couples to amino acid residues between K310 and K318.
Expression of the chicken angiotensin II receptor in adrenal
Aves have a unique angiotensin II receptor, whose character is different from AT1 and AT2. The probe used for the hybridization to determine the expression of the receptor was synthesized using PCR.The receptor was expressed in adrenal but not in liver.
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