| Project/Area Number |
09671904
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional basic dentistry
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| Research Institution | Meikai University |
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Principal Investigator |
TOMOMURA Akito Meikai University, Sch.of Dentistry Dept.of Biochemistry, Assistant Professor, 歯学部, 講師 (60188810)
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| Co-Investigator(Kenkyū-buntansha) |
INABA Akemi Meikai University, Sch.of Dentistry Dept.of Biochemistry, Research Associate, 歯学部, 助手 (50151585)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Osteoclast / Bone resorption / Protease / Receptor / カルシウム |
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| Research Abstract |
A reduction of serum calcium (hypocalcemia) occurs in patients with acute pancreatitis. We purified the serum calcium-decreasing factor from pancreas and named caldecrin, which found to be a chymotrypsin-type serine protease. Science the caldecrin suppressed a bone resorption, we studied to determine a suppressive domains for the bone resorption and its receptors. Caldecnn fragments were fused to Hislidine-tagged thioredoxin to produce disulfide bridge. Expression vector with caldecrin fragment were transformed into E.Coli, induced with IPTG, and purified by affinity chelating column. Bone resorption activity was measured by a method of excavated bone resorption pit formation by rabbit osteoclasts. The caldecrin is composed of 2 beta-barrel like structures. When the caldecrin was divided into 2 parts, the suppression activities for bone resorption were found to be both parts. The suppressive activity was localized on a latter half part of the former barrel structure. Disruption of disulFide bridge of the latter barrel structure was not changed the suppressive activity. Caldecrin suppressed the bone resorption of purified rabbit osteoclasts, suggesting the existence of caldecrin receptors on the rabbit osteoclasts. Thus, we made rabbit osteoclast cDNA library from lO^4 osteoclasts using PCR with SMART oligo and oligo (dT) primers. Recombinant E.Colis with expression vector were divided into subpools. Thus we could established the cloning system for rabbit osteoclst receptor genes using anti-thioredoxin antibody.
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