Recombinant Kinase Production and Fragment Screening by NMR Spectroscopy.
During the past decade fragment-based drug discovery (FBDD) has rapidly evolved and several drugs or drug candidates developed by FBDD approach are clinically in use or in clinical trials. For example, vemurafenib, a V600E mutated BRAF inhibitor, was developed by utilizing FBDD approach and approved by FDA in 2011. In FBDD, screening of fragments is the starting step for identification of hits and lead generation. Fragment screening usually relies on biophysical techniques by which the protein-bound small molecules can be detected. NMR spectroscopy has been extensively used to study the molecular interaction between the protein and the ligand, and has many advantages in fragment screening over other biophysical techniques. This chapter describes the practical aspects of fragment screening by saturation transfer difference NMR.
Department of Pharmacy, Dongguk University-Seoul, 32 Dongguk-ro, Ilsandong-gu, Goyang, Gyeonggi, 410-820, Republic of Korea.
Byeonggu
B
Han
Department of Pharmacy, Dongguk University-Seoul, 32 Dongguk-ro, Ilsandong-gu, Goyang, Gyeonggi, 410-820, Republic of Korea. hcahn@dongguk.edu.
Hee-Chul
HC
Ahn
D003001:Cloning, Molecular / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D055808:Drug Discovery / Q000379:methods; D004926:Escherichia coli; D006801:Humans; D007202:Indicators and Reagents; D008024:Ligands; D048057:Mitogen-Activated Protein Kinase 10 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010446:Peptide Fragments / Q000032:analysis*; D019151:Peptide Library; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D011494:Protein Kinases / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D020349:Surface Plasmon Resonance / Q000379:methods
D007202:Indicators and Reagents; D008024:Ligands; D010446:Peptide Fragments; D019151:Peptide Library; D047428:Protein Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; D048057:Mitogen-Activated Protein Kinase 10
Fragment; Fragment-based drug discovery; Kinase; NMR screening; Optimum solubility screening; Saturation transfer difference; Surface plasmon resonance; X-ray crystallography