1990 Fiscal Year Final Research Report Summary
Purification of a Photoreceptor-Specific Meka Protein and its Physiological Function
| Project/Area Number |
01571254
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
応用薬理学・医療系薬学
|
|---|
| Research Institution | Kanazawa University |
|---|
Principal Investigator |
KUO Cheーhui Cancer Research Institute Research Associate, がん研究所, 助手 (50126570)
|
|---|
| Project Period (FY) |
1989 – 1990
|
| Keywords | MEKA cDNA / MEKA protein / Photoreceptor cell / Transducin / G protein / Retina |
|---|
| Research Abstract |
MEKA cDNA isolated from bovine retina cDNA library encodes acidic soluble (MEKA) protein whose molecular weight is estimated to be about 30,000. From the MEKA cDNA ligated with expression vector, MEKA fusion protein could be synthesized in E. Coli. It was used as an antigen to prepare an anti-MEKA serum.
(1) Human MEKA genome was isolated by the MEKA cDNAas a prove. It consists of two introns and three exons. There is about 90% homology of amino acid sequence between bovine and human MEKA proteins.
(2) A Immunohistochemical study with anti-MEKA revealed that MEKA protein is not only expressed in a rod photoreceptor cell, but also in a cone photoreceptor cell. Translocation of the MEKA protein in a photoreceptor cell occurred by light stimulation. Purification of the MEKA protein by using the anti-MEKA from bovine retina was carried out. The MEKA protein (32 KDa) forms a trimeric complex (74 KDa) with (36 KDa) and (10 KDa) subunits of transducin G protein.
(3) A MEKA homologous (BL-MEKA) cDNA clone was isolated from bovine a liver cDNA library and analyzed. I found that nucleotide sequence of the BL-MEKA cDNA shows high homology to retinal MEKA cDNA.
|
