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• W3205346021 endingPage "131" @default.
• W3205346021 startingPage "65" @default.
• W3205346021 abstract "NMR spectroscopy has proven itself to be invaluable in probing the structures of proteins that are not amenable to crystallization. In addition, NMR spectroscopy is of great value in characterizing the weak interactions that form the basis of drug-like action of potential hits. While 2D NMR methods for the characterization of small molecules were developed in the 1970s, in recent times, there has been a significant growth in the macromolecular NMR field, which has potentiated structure determination of several drug target proteins and protein–ligand complexes and discovery of inhibitors through NMR-based screening methods. In this chapter, our aim will be to describe the state-of-the-art methods adopted for isotope labeling of proteins, which will be followed by description of the multidimensional NMR experiments and their utilization for determination of solution structure of the proteins. NMR-based structure determination begins with suitable isotope labeling of proteins using many innovative methods. Once a double or triple isotopically labeled sample has been made, a series of multidimensional NMR experiments are carried out for the assignment of chemical shifts of the backbone and side-chain resonances and for measuring the NOE between identified protons. An ensemble of structures can then be calculated by incorporating the experimentally derived dihedral angle and distance constraints. Refinement of structural quality is done through additional constraints derived from Residual Dipolar Couplings and Paramagnetic Relaxation Enhancements. Using this technique and protocols, structures of several drug target proteins have been successfully determined with high precision. Further, NMR has been used to screen the small molecules that bind to the target protein and to study the structural and dynamics aspects of protein-ligand complexes." @default.
• W3205346021 created "2021-10-25" @default.
• W3205346021 creator A5023995827 @default.
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• W3205346021 creator A5077711991 @default.
• W3205346021 date "2021-01-01" @default.
• W3205346021 modified "2023-09-27" @default.
• W3205346021 title "Protein Labeling and Structure Determination by NMR Spectroscopy" @default.
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