A protein recycling system for nuclear magnetic resonance-based screening of drug candidates.
A sample-treating system for nuclear magnetic resonance (NMR)-based interaction screening between drug candidates (small molecules) and a protein of interest was developed by applying high-performance liquid chromatography (HPLC) technology. The system prepares a test solution by mixing a (15)N-labeled protein solution and a solution of each candidate compound, loads it to a flow cell-type NMR probe, and recycles the protein after the data acquisition. The system was designed to behave differently according to the information obtained in NMR measurements. In a test operation with a 100-compound library, the system could single out known interacting substances properly. Recovery values of the protein and one representative compound were 75 and 71%, respectively, and the recovered protein was found intact as intended.
Hirayama
Aya
A
Shiseido Material Science Research Center, Tsuzuki-ku, Yokohama, Kanagawa, Japan.
Shirota
Osamu
O
Nomura
Mitsuru
M
Nagadoi
Aritaka
A
Nishimura
Yoshifumi
Y
D002474:Cell-Free System; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015195:Drug Design; D006859:Hydrogen / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009211:Myoglobin / Q000032:analysis; D009590:Nitrogen Radioisotopes / Q000737:chemistry; D004364:Pharmaceutical Preparations / Q000032:analysis*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D011868:Radioisotopes / Q000737:chemistry; D012107:Research Design; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry
D009211:Myoglobin; D009590:Nitrogen Radioisotopes; D004364:Pharmaceutical Preparations; D011506:Proteins; D011868:Radioisotopes; D006859:Hydrogen; D013762:Tetrahydrofolate Dehydrogenase