NMR Methodology For Detection Of Hydrogen Bonds In Proteins And Protein Complex Interfaces

Nuclear Magnetization Resonance is playing more and more important role in study of protein structure,dynamics and interaction which benefits from the development of the technique on NMR hardware and experiment.This thesis focuses on the new NMR experimental development and its application of protein study, including hydrogen bond detection and protein complex interface mapping by NMR method.In chapter one,a brief introduction of protein NMR is provided.We also present some development of NMR technique:TROSY and CRINEPT,Residue Dipolar Coupling(RDC)and Reduced Dimension(RD)NMR.Chapter two provides an exploration of hydrogen bond detection in protein through heteronuclear NOESY.Hydrogen bonds in protein play important role in protein structures.There have been some NMR methods for detection of hydrogen bonds.The development of 2D heteronuclear NOESY(HOESY)is reviewed and a rotating frame HOESY has been designed for detection of NOE correlation in large molecules.Meanwhile,we present a new 2D ¹³C-¹H HOESY with presaturation and a significant sensitivity enhancement is obtained.In chapter three we present a new NMR method for mapping protein-ligand interaction which is named as Water Accessibility Perturbation(WAP).Study of protein-ligand interaction has a great significance on biological function.NMR is well suited for study of protein-ligand interaction.SEA-HSQC experiments can study the chemical exchange between the amide protons and water molecules.The difference of the exchange rate after binding can be used to mapping protein-ligand interaction.The method has been tested on the Hox-Gem and CaM-M13 complexes and obtains good results.