2002 Fiscal Year Final Research Report Summary
Protein Microarrays based on C-terminal Labeling of Proteins
| Project/Area Number |
13559007
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
広領域
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| Research Institution | Keio University |
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Principal Investigator |
YANAGAWA Hiroishi Fac. Sci. & Tech., Professor, 理工学部, 教授 (40327672)
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| Co-Investigator(Kenkyū-buntansha) |
DOI Nobuhide Fac. Sci. & Tech., Assistant Professor, 理工学部, 講師 (50327673)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Proteomics / Proteome / Micro array / Protein Chip / Fluorescence Labeling / Interaction / Puromycin |
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| Research Abstract |
Protein microarrays or proteome chips are potentially powerful tools for comprehensive analysis of protein-protein interactions. In interaction analysis, a set of immobilized proteins is arrayed on slides and each slide is probed with a set of fluorescently labeled proteins. Here we have developed and tested an 'in vitro' protein microarray, in which both arraying and probing proteins were prepared by cell-free translation. The in vitro synthesis of fluorescently labeled proteins was accomplished by a new method: a fluorophore-puromycin conjugate was incorporated into a protein at the C-terminus on the ribosome. The resulting fluorescently labeled proteins were confirmed to be useful for probing protein-protein interactions on protein microarrays in model experiments. We have used three pairs of model proteins that are known to interact: the classical oncogenes, c-Fos and c-Jun; Polycomb-related genes, RYBP and Ring1A; and calmodulin binding peptide (CBP) derived from skeletal muscle myosin light-chain kinase and calmodulin. The in vitro protein microarrays can easily be extended to a high-throughput format and thus should be useful for large-scale analysis of protein-protein interactions.
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[Publications] Kawahashi, Y., Doi, N., Takashima, H., Tsuda, C., Oishi, Y., Oyama, R., Yonezawa, M., Miyamoto-Sato, E., Yanagawa, H.: "In vitro protein microarrays for detecting protein-protein interactions: Application of a new method for fluorescence labeling of proteins"Proteomics. in press. (2003)
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「研究成果報告書概要(欧文)」より
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[Publications] Doi, N, Takashima, H., Kinjo, M., Sakata, K., Kawahashi, Y., Oishi, Y., Oyama, R., Miyamoto-Sato, E., Sawasaki, T., Endo,Y., Yanagawa, H.: "Novel fluorescence labeling and high-throughput assay technologies for in vitro analysis of protein interactions"Genome Research. 12. 487-492 (2002)
Description
「研究成果報告書概要(欧文)」より
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