2014 Fiscal Year Final Research Report
Preparation of monoclonal antibody against sulfated protein and its application to sulfo-proteomics studies
| Project/Area Number |
23510265
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Living organism molecular science
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| Research Institution | Nigata University of Phermacy and Applied Life Sciences |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
ASADA Shinichi 新潟薬科大学, 薬学部, 助教 (50424883)
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| Project Period (FY) |
2011-04-28 – 2015-03-31
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| Keywords | 硫酸化タンパク質 / モノクローナル抗体 / ファージディスプレイ / チロシン硫酸化酵素 / 基質配列モチーフ / ペプチド合成 / プロテオーム解析 |
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| Outline of Final Research Achievements |
We have aimed to obtain the monoclonal antibody against sulfated tyrosine as immuno-chemical tool applicable to the sulfo-proteomics studies; however specific antibody against sulfated proteins could not be raised by the conventional immunization protocol in mice due to the inherent instability of sulfate. Thus we changed the approach to the phage-display method and now fundamental studies are in progress.
We have established the expression system in E. coli for two enzymes (TPST-1 and -2) responsible for tyrosine sulfation in protein. Using obtained proteins having enzyme activity and synthetic peptides based on ADYAE sequence, we found out the XDY (X: various amino acids) motif as a minimal peptide substrate for sulfation. We have developed a preparation procedure for peptide thioester containing tyrosine sulfate residue, and using the Ag+-mediated thioester segment condensation, we obtained the long-chain peptides containing multiple tyrosine sulfate residues in high yields.
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| Free Research Field |
ペプチド・タンパク質化学
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