Development of a chemical screening system using aqueorin-fused protein. 2008

Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.

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.

UI MeSH Term Description Entries
D008163 Luminescent Measurements Techniques used for determining the values of photometric parameters of light resulting from LUMINESCENCE. Bioluminescence Measurements,Bioluminescent Assays,Bioluminescent Measurements,Chemiluminescence Measurements,Chemiluminescent Assays,Chemiluminescent Measurements,Chemoluminescence Measurements,Luminescence Measurements,Luminescent Assays,Luminescent Techniques,Phosphorescence Measurements,Phosphorescent Assays,Phosphorescent Measurements,Assay, Bioluminescent,Assay, Chemiluminescent,Assay, Luminescent,Assay, Phosphorescent,Assays, Bioluminescent,Assays, Chemiluminescent,Assays, Luminescent,Assays, Phosphorescent,Bioluminescence Measurement,Bioluminescent Assay,Bioluminescent Measurement,Chemiluminescence Measurement,Chemiluminescent Assay,Chemiluminescent Measurement,Chemoluminescence Measurement,Luminescence Measurement,Luminescent Assay,Luminescent Measurement,Luminescent Technique,Measurement, Bioluminescence,Measurement, Bioluminescent,Measurement, Chemiluminescence,Measurement, Chemiluminescent,Measurement, Chemoluminescence,Measurement, Luminescence,Measurement, Luminescent,Measurement, Phosphorescence,Measurement, Phosphorescent,Measurements, Bioluminescence,Measurements, Bioluminescent,Measurements, Chemiluminescence,Measurements, Chemiluminescent,Measurements, Chemoluminescence,Measurements, Luminescence,Measurements, Luminescent,Measurements, Phosphorescence,Measurements, Phosphorescent,Phosphorescence Measurement,Phosphorescent Assay,Phosphorescent Measurement,Technique, Luminescent,Techniques, Luminescent
D008727 Methotrexate An antineoplastic antimetabolite with immunosuppressant properties. It is an inhibitor of TETRAHYDROFOLATE DEHYDROGENASE and prevents the formation of tetrahydrofolate, necessary for synthesis of thymidylate, an essential component of DNA. Amethopterin,Methotrexate Hydrate,Methotrexate Sodium,Methotrexate, (D)-Isomer,Methotrexate, (DL)-Isomer,Methotrexate, Dicesium Salt,Methotrexate, Disodium Salt,Methotrexate, Sodium Salt,Mexate,Dicesium Salt Methotrexate,Hydrate, Methotrexate,Sodium, Methotrexate
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D000331 Aequorin A photoprotein isolated from the bioluminescent jellyfish Aequorea. It emits visible light by an intramolecular reaction when a trace amount of calcium ion is added. The light-emitting moiety in the bioluminescence reaction is believed to be 2-amino-3-benzyl-5-(p-hydroxyphenyl)pyrazine (AF-350). Aequorine
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D013762 Tetrahydrofolate Dehydrogenase An enzyme of the oxidoreductase class that catalyzes the reaction 7,8-dihyrofolate and NADPH to yield 5,6,7,8-tetrahydrofolate and NADPH+, producing reduced folate for amino acid metabolism, purine ring synthesis, and the formation of deoxythymidine monophosphate. Methotrexate and other folic acid antagonists used as chemotherapeutic drugs act by inhibiting this enzyme. (Dorland, 27th ed) EC 1.5.1.3. Dihydrofolate Dehydrogenase,Dihydrofolate Reductase,Folic Acid Reductase,Acid Reductase, Folic,Dehydrogenase, Dihydrofolate,Dehydrogenase, Tetrahydrofolate,Reductase, Dihydrofolate,Reductase, Folic Acid
D016503 Drug Delivery Systems Systems for the delivery of drugs to target sites of pharmacological actions. Technologies employed include those concerning drug preparation, route of administration, site targeting, metabolism, and toxicity. Drug Targeting,Delivery System, Drug,Delivery Systems, Drug,Drug Delivery System,Drug Targetings,System, Drug Delivery,Systems, Drug Delivery,Targeting, Drug,Targetings, Drug
D018189 Immunomagnetic Separation A cell-separation technique where magnetizable microspheres or beads are first coated with monoclonal antibody, allowed to search and bind to target cells, and are then selectively removed when passed through a magnetic field. Among other applications, the technique is commonly used to remove tumor cells from the marrow (BONE MARROW PURGING) of patients who are to undergo autologous bone marrow transplantation. Immunomagnetic Bead Technique,Immunomagnetic Purging,Immunomagnetic Cell Separation,Bead Technique, Immunomagnetic,Bead Techniques, Immunomagnetic,Cell Separation, Immunomagnetic,Cell Separations, Immunomagnetic,Immunomagnetic Bead Techniques,Immunomagnetic Cell Separations,Immunomagnetic Purgings,Immunomagnetic Separations,Purging, Immunomagnetic,Purgings, Immunomagnetic,Separation, Immunomagnetic,Separation, Immunomagnetic Cell,Separations, Immunomagnetic,Separations, Immunomagnetic Cell
D025941 Protein Interaction Mapping Methods for determining interaction between PROTEINS. Interaction Mapping, Protein,Interaction Mappings, Protein,Mapping, Protein Interaction,Mappings, Protein Interaction,Protein Interaction Mappings

Related Publications

Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
August 2015, Bioorganic chemistry,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
January 2011, Bioscience, biotechnology, and biochemistry,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
September 1993, The Journal of biological chemistry,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
September 1982, Journal - Association of Official Analytical Chemists,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
July 2017, Biosensors & bioelectronics,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
April 2019, Sheng wu gong cheng xue bao = Chinese journal of biotechnology,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
February 2002, Molecules and cells,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
June 2012, RNA (New York, N.Y.),
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
December 2019, Cell chemical biology,
Masafumi Suzuki, and Shoichiro Furukawa, and Chikanori Kuramori, and Chika Sawa, and Yasuaki Kabe, and Mitsuhiro Nakamura, and Jun-Ichi Sawada, and Yuki Yamaguchi, and Satoshi Sakamoto, and Satoshi Inouye, and Hiroshi Handa
February 2020, Molecules (Basel, Switzerland),
Copied contents to your clipboard!