2000 Fiscal Year Final Research Report Summary
Development of fine micro analytical method of drug recognition function of sugar chains
| Project/Area Number |
11307054
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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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 | KYOTO UNIVERSITY |
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Principal Investigator |
NAKAGAWA Terumichi Kyoto University, Graduate School of Pharmaceutical Sciences, Professor, 薬学研究科, 教授 (70025708)
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| Co-Investigator(Kenkyū-buntansha) |
SHIBUKAWA Akimasa Kyoto University, Graduate School of Pharmaceutical Sciences, Associate Professor, 薬学研究科, 助教授 (30170913)
KURODA Yoshihiro Kyoto University, Graduate School of Pharmaceutical Sciences, Associate Professor, 薬学研究科, 助教授 (90093236)
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| Project Period (FY) |
1999 – 2000
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| Keywords | glycochain / plasma protein binding / microheterogeneity / high-performance frontal analysis / ESI-T0F-MS / sodium channel inactivation gate peptide / prion protein / capillary electrophoresis |
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| Research Abstract |
A novel analytical system using capillary electrophoresis (CE) and mass spectroscopy (MS) was developed to investigate the structural effect of glycoproteins upon drug binding affinity. α1-acid glycoprotein (AGP) was used as a model glycoprotein, and the binding affinities of AGP glycoforms and genetic variants to model basic drugs were evaluated.
(a) The biantennary glycochains which locate only near the drug binding site on AGP molecule does not give any significant effect upon the enantioselective binding of AGP to model basic drugs (disopyramide and verapamil).
(b) AGP has three major genetic variants, A-variant, F1-variant and S-variant. A-variant shows higher enantioselectivity in the binding with the enantiomers of model drugs than F1^*S variants, while these genetic variants show almost the same enantioselectivity in the binding to verapamil enantiomers.
(c) CE/MS system using sheathless ESI method allows one-order higher detectability in the analyses of tryptic digests of AGP var
… More
iants than conventional CE method using UV detection.
In order to investigate the effect of microheterogeneity of glycoproteins upon the function and the secondary structure of the proteins, the structural chemistry of several synthesized peptide fragments of Na-channel protein and prion protein were studied by using NMR, molecular orbital method and CE spectroscopy. The synthesized fragments correspond to the major peptide portion responsible for the secondary structure and/or the function of the proteins. The followings were found.
(a) The secondary structure of model peptide fragments changes depending on the hydrophobicity of environment due to the nature and the component of solvents.
(b) Glycans containing both hydrophobic moiety and hydrophobic moiety create local hydrophobic space with different micro-permittivity depending on nature, sequence and the branching structure of glycans. It is suggested that the creation of hydrophobic local environment would cause the change of secondary structure of protein, resulting in the regulation of function and amyloid fibril formation. Less
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Research Products
(16 results)
