Cloning of a kallikrein-like enzyme originated from dog heart and investigation of its intracellular localization and expression regulation.
| Project/Area Number |
10670692
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Circulatory organs internal medicine
|
|---|
| Research Institution | Fukuoka University |
|---|
Principal Investigator |
SASAGURI Manabu Fukuoka University. School of Medicine. Lecturer, 医学部, 講師 (00178675)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NODA Keita Fukuoka University. School of Medicine. Lecturer, 医学部, 講師 (70289536)
|
|---|
| Project Period (FY) |
1998 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,500,000 (Direct Cost: ¥1,500,000)
|
|---|
| Keywords | kallikrein / kinin / angiotensin / kinin-tensin-system |
|---|
| Research Abstract |
We have previously shown that the purified kinin-forming enzyme from the dog heart is similar to tissue kallikrein and is also able to convert angiotensin (Ang) I to Ang II. The aim of the present study is to clarify the cloning, intracellular localization and expression regulation of this enzyme. Western blot analysis indicated the presence of this enzyme not only the heart (atria, ventricle, coronary artery) but also other tissues such as aorta, kidney, pancreas, small intestine, and skeletal muscle. Its presence was also demonstrated in MDCK cell originated from canine distal tubules. The kallikrein-like enzyme has been purified from homogenates of the kidney and aorta by a DEAE-Sepharose and aprotinin affinity column. Purified enzyme from the kidney and aorta was visualized at the relative molecular weight of 65 kDa on Western blotting which is identical to that of the kallikrein-like enzyme purified from the heart. The purified enzyme from the heart was proceeded to amino acid composition and sequence analysis. It has N-terminal blocked amino acid. The amino acid composition of purified kallikrein-like enzyme from the heart was different from that of dog urinary and pancreatic kallikrein. Autolysed samples of this enzyme were electroblotted to the membrane, and cutted membrane including one band was proceeded to amino acid sequence analysis. Three sets of partial amino acid sequence were obtained. These sequences are different to the hitherto known kallikrein. Mixed primer was newly made, and cDNA library from the dog heart was also newly prepared. At the present, DNA sequence analysis is now ongoing. This enzyme may play an important role in cardiovascular systems through generating kinins and Ang II.
|
Report
(3 results)
Research Products
(9 results)
