Document Detail

Development of a chemical screening system using aqueorin-fused protein.
MedLine Citation:
PMID:  18243129     Owner:  NLM     Status:  MEDLINE    
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.
Masafumi Suzuki; Shoichiro Furukawa; Chikanori Kuramori; Chika Sawa; Yasuaki Kabe; Mitsuhiro Nakamura; Jun-Ichi Sawada; Yuki Yamaguchi; Satoshi Sakamoto; Satoshi Inouye; Hiroshi Handa
Related Documents :
12442549 - Prevention of developmental abnormalities with particular emphasis of primary prevention.
20356939 - Distribution of nuchal translucency in antenatal screening for down's syndrome.
22495259 - The vascular endothelial growth factor family in adverse pregnancy outcomes.
12378579 - Second trimester trisomy 21 maternal serum marker screening. results of a countrywide s...
17851819 - Maternal anxiety about first trimester nuchal translucency screening and impact of posi...
12768409 - Impact of public health strategies on the birth prevalence of cystic fibrosis in britta...
9292639 - Pregnancy-associated deaths in finland 1987-1994--definition problems and benefits of r...
19777489 - Dependence of maternal serum [afp]/[hcg] median ratios on age of gestation: comparison ...
15035549 - Involvement of interleukin-1 and the interleukin-1 receptor antagonist in in vitro embr...
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    
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
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
Reg. No./Substance:
50934-79-7/Aequorin; 59-05-2/Methotrexate; EC Dehydrogenase

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

Previous Document:  Perilipin, a critical regulator of fat storage and breakdown, is a target gene of estrogen receptor-...
Next Document:  Inhibitory crosstalk between ERK and AMPK in the growth and proliferation of cardiac fibroblasts.