| Project/Area Number |
14540561
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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 |
分離・精製・検出法
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| Research Institution | Ehime University |
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Principal Investigator |
MANABE Takashi Ehime University(Faculty of Science), Dept.Chemistry, Professor, 理学部, 教授 (60094281)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMAZAKI Yoji Ehime University(Faculty of Science), Dept.Chemistry, Instructor, 理学部, 助手 (80284389)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | plasma proteins / two-dimensional electrophoresis / isoelectric focusing / SDS-gel electrophoresis / minor proteins / mass spectrometry / MALDI-TOP MS / ESI-MS / MS / MALDI-TOF MS / 血漿 / タンパク質 / 電気泳動 / 微量 / 分離 / 構造解析 |
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| Research Abstract |
We have examined the origin of human plasma minor proteins which we found comparing the patterns of two-dimensional gel electrophoresis run under two different conditions of size-separation (second dimension electrophoresis) ; one in the absence of denaturing agents throughout the run and one in the presence of SDS during the size-separation electrophoresis. The minor proteins were analyzed by mass spectrometry, identification of proteins by peptide mass fingerprinting using matrix-assisted laser desorption ionization-time-of-flignt mass spectrometer (MALDI-TOF MS) and partial amino acid sequencing by mass-tag method using electrospray ionization tandem mass spectrometer (ESI-MS/MS). Further, we have developed a three-step electrophoresis technique which employ slab-agarose gels in the step of isoelectric focusing and showed that this is useful for the structural analysis of the minor proteins. The comparisons of the patterns obtained by using different conditions of two-dimensional electrophoresis, together with the structural analyses of the proteins on the patterns, will provide insights on the construction processes of functional protein complexes.
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