Document Detail


Development of a chemical screening system using aqueorin-fused protein.
MedLine Citation:
PMID:  18243129     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We developed a unique screening system that consists of combination of high photo-sensitivity of photoprotein aequorin (AQ) and our developed high-performance affinity purification system. In the present study, we demonstrated to detect the specific interaction between methotrexate (MTX) and its target dihydrofolate reductase (DHFR) fused with AQ. We succeeded to prepare highly purified AQ-fused DHFR, which showed high sensitive light emission. To test the screening system, we prepared the complex of MTX-immobilized magnetic nanobeads and AQ-fused DHFR. Bound AQ-fused DHFR with the beads was specifically released by addition of MTX. Thus, this methodology enables us to search a novel chemical that binds to target proteins without complicated processes. Furthermore, thank to the highly sensitive luminescence intensity of AQ, this methodology would be performed in very small scale with high responsibility, leading to development of high throughput screening systems.
Authors:
Masafumi Suzuki; Shoichiro Furukawa; Chikanori Kuramori; Chika Sawa; Yasuaki Kabe; Mitsuhiro Nakamura; Jun-Ichi Sawada; Yuki Yamaguchi; Satoshi Sakamoto; Satoshi Inouye; Hiroshi Handa
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-02-01
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  368     ISSN:  1090-2104     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2008 Apr 
Date Detail:
Created Date:  2008-03-04     Completed Date:  2008-04-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  600-5     Citation Subset:  IM    
Affiliation:
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
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MeSH Terms
Descriptor/Qualifier:
Aequorin / analysis*
Drug Delivery Systems / methods
Immunomagnetic Separation / methods*
Luminescent Measurements / methods*
Methotrexate / analysis*,  chemistry
Protein Binding
Protein Interaction Mapping / methods*
Sensitivity and Specificity
Tetrahydrofolate Dehydrogenase / analysis*,  chemistry
Chemical
Reg. No./Substance:
50934-79-7/Aequorin; 59-05-2/Methotrexate; EC 1.5.1.3/Tetrahydrofolate Dehydrogenase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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