| Project/Area Number |
06671588
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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 |
Urology
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| Research Institution | Osaka University |
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Principal Investigator |
YOSHIOKA Toshiaki Osaka University, Dept.of Urology, Associate Professor, 医学部, 講師 (30191547)
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| Co-Investigator(Kenkyū-buntansha) |
HONDA Masahito Osaka University, Dept.of Urology, Assistant Professor, 医学部, 助手 (70263291)
KOKADO Yukito Osaka University, Dept.of Urology, Associate Professor, 医学部, 講師 (30186639)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Calcium oxalate crystal / Urinary macromolecules / Urinary stone / Urinary protein / 高分子物質 |
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| Research Abstract |
Urinary macromolecules are considered to play a important role in stone formation. The aim of our study is to characterize the urinary protein affecting stone formation.
1.We obtained crystal surface binding substance (CSBS) by adding calcium and oxalate to human whole urine. The CSBS is urinary macromolecules absorbed onto the surface of calcium oxalate crystals and yields strong inhibitory activity on calcium oxalate crystallization. Proteins contained in the CSBS are analyzed.
2.Fractions enriched in proteins with strong calcium oxalate crystal growth inhibitory activity are obtained by DEAE-Sepharose CL-6B ion-chromatography. These fractions are fractionated by hydroxyapatite column with stepwise elution of sodium phosphate.
3.The SDS-PAGE shows many bands of protein. Two major proteins, molecular weight of 30Kd and 67Kd, are analyzed by a amino acid sequencer. As a result, 30Kd protein is identified with human prothrombin and 67Kd protein with human osteopontin.
4.Protein composition of fractions obtained by hydroxyapatite column is also analyzed by immunoblotting. Human serum albumin, alpha 1-acid glycoprotein, alpha 1-microglobulin, alpha 2-HS glycoprotein, retinol-binding protein, transferrin, Tamm-Horsfall mucoprotein and prothrombin are detected. However, it is considered that these proteins does not play a signifficant role on calcium oxalate crystal formation because each fractions containing each proteins reveal weak inhibitory power on crystal growth.
We conclude that other unknown proteins included in CSBS are thought to be dominant protein inhibitors.
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