| Project/Area Number |
13672267
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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 |
Physical pharmacy
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| Research Institution | Mukogawa Women's University |
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Principal Investigator |
MATSUNAGA Hisami Mukogawa Women's University, Faculty of Pharmaceutical Sciences, Lecturer, 薬学部, 講師 (70271418)
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| Co-Investigator(Kenkyū-buntansha) |
HAGINAKA Jun Mukogawa Women's University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (20164759)
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| Project Period (FY) |
2001 – 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: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | chicken α_1-acid glycoprotein / MALDI-TOF MS / disulfide bond / sugar chain / 糖タンパク質 / 固定化カラム / 光学分割 / 光学認識機構 / ジスルフィド結合 / アミノ酸配列 / 医薬品活性体 / 糖タンパク質の遺伝子組み換え |
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| Research Abstract |
Recently, we reported the amino acid sequence of chicken α_1-acid glycoprotein(chicken α_1-AGP)(Biochem.Biophys.Res.Commun.295(2002)587-590). In this study, we identify disulfide bonds and site-specific glycosylation in chicken α_1-AGP using tryptic digests of carbamidomethylated, chicken α_1-AGP, carbamidomethylated, completely deglycosylated chicken α_1-AGP(cd-α_1-AGP), and non-reduced, denatured cd-α_1-AGP by matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS). Based on the existence of the peptides detected at m/z 3037.4(amino acid sequence 69-76 plus 161-183) and 3453.3(amino acid sequence 69-80 plus 161-183), the location of two disulfide bonds of chicken α_1-AGP was determined to be Cys 6-Cys 146 and Cys 73-Cys 163. Furthermore, it was found that Asn 16,70,77 and 87 were fully glycosylated, and that Asn 62 was partially glycosylated.
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