An Efficient Protocol for NMR-Spectroscopy-Based Structure Determination of Protein Complexes in Solution

摘要

Eukaryotic proteins typically have a modular architecture, characterized by multiple structural domains that are connected by flexible linkers. Regulation of cellular processes depends on an interaction network between these individual modules and the formation of a quaternary structure. Dynamic rearrangement, often coupled to ligand binding, is a common feature of these multicomponent systems. While compact and rigid complexes can be efficiently studied using X-ray crystallography, protein complexes or multidomain proteins that involve weak and transient domain interactions should be preferably investigated using solution techniques. A number of NMR spectroscopy studies of protein complexes have been reported in recent years. However, a general protocol is not available, and many applications still rely on NOE-based interdomain distance restraints, which are difficult to obtain and to assign in high-molecular-weight systems.