| Project/Area Number |
62570844
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Functional basic dentistry
|
|---|
| Research Institution | The Nippon Dental University |
|---|
Principal Investigator |
SHIMOMURA Hiromi The Nippon Dental Univ., Niigata, Oral Biochemistry, Associate Prof., 新潟歯学部, 助教授 (40139259)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TERADA Akane The Nippon Dental Univ., Niigata, Oral Biochemistry, Researcher, 新潟歯学部, 助手 (60180080)
|
|---|
| Project Period (FY) |
1987 – 1989
|
| Project Status |
Completed (Fiscal Year 1989)
|
| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1988: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| Keywords | parotid gland / submandibular gland / protein kinase C / substrate / adrenergic receptor / gene structure / β_1-アドレナリン受容体 / ラット / 遺伝子 / 構造 / cDNA / Lambdc charon35 / プロテイン、キナーゼ / C / α_1-アドレナリン受容体 / H-7 / プロティンカイネースC / アミラーゼ / phorbol ester |
|---|
| Research Abstract |
1)Role of protein kinase C on amylase secretion--a)The activities of Ca^2 /phospholipid- dependent protein kinase(protein kinase C) and Ca^2 /calmodulin-dependent protein kinase II(protein kinase II)in rat salivary gland were assayed using synthetic peptide syntide-2(Pro-Leu-Ala-Arg-Thr-Leu-Ser-Val-Ala-Gly-Leu-Pro-Gly-Lys-Lys) as substrate. Levels of the protein kinase C and protein kinase II were less than 0.05 units/g in the parotid and submandibular glands. The protein kinase C inhibitor, H-7, inhibited amylase secretion from rat parotid gland stimulated by 12-O-tetradecanoylphorbol 13- acetate(PMA) or the combination of phosphatidylserine and 1,2-diolein. The results supported the hypothesis of the secretory mechanism that protein kinase C mediates amylase secreiton in rat parotid gland. b) Phosphorylation by protein kinase C in salivary gland were investigated. When the homogenates of rat parotid and submandibular glands were subjected to phosphorylation with the addition of extra
… More
cellular protein kinase C, the major Mr. 8OK-Da band(SDS-PAGE) appeared. This band was disappeared by the addition of protein kinase C inhibitor, H-7. The Mr.80k-Da substrate was localized predominantly in the 100,000g supernatant fraction on the case of parotid gland. In the case of submandibular gland, the substrate(80-KDa protein) appeared to be approximately equally distributed between the 100,000g supernatant fraction(cytosolic fraction) and the 100,000g pellet fraction(granulate fraction).
2)Cloning of rat beta_1-adrenergic receptor gene--- A rat heart cDNA library( Clontech, USA) and rat genomic DNA library(prepared from Eco RI digests of rat genomic DNA and Lambda charon 35 Eco RI vector) were screened with the synthetic oligonucleotide based on human beta_1-AR gene. Three clones from cDNA and one clone from genomic library were isolated. Sequencing analysis of these clones revealed that rat beta_1-AR gene is intronless. The amino acid sequence of rat beta_1-AR is 90% identical to that of the human beta_1-AR. Less
|
