Nmr screening for rapid protein characterization in structural proteomics

Abstract

In the age of structural proteomics when protein structures are targeted on a genome-wide scale, the identification of proteins that are amenable to analysis using x-ray crystallography or NMR spectroscopy is the key to high throughput structure determination. NMR screening is a beneficial part of a structural proteomics pipeline because of its ability to provide detailed biophysical information about the protein targets under investigation at an early stage of the structure determination process. This chapter describes efficient methods for the production of uniformly15N-labeled proteins for NMR screening using both conventional IPTG induction and autoinduction approaches in E. coli. Details of sample preparation for NMR and the acquisition of 1D1H NMR and 2D1H-15N HSQC spectra to assess the structural characteristics and suitability of proteins for further structural studies are also provided.