Chemistry Reference
In-Depth Information
Contemporary Methods in Structure
Determination of Membrane Proteins
by Solution NMR
Tabussom Qureshi and Natalie K. Goto
Abstract Integral membrane proteins are vital to life, being responsible for infor-
mation and material exchange between a cell and its environment. Although high-
resolution structural information is needed to understand how these functions are
achieved, membrane proteins remain an under-represented subset of the protein
structure databank. Solution NMR is increasingly demonstrating its ability to help
address this knowledge shortfall, with the development of a diverse array of
techniques to counter the challenges presented by membrane proteins. Here we
document the advances that are helping to define solution NMR as an effective
tool for membrane protein structure determination. Developments introduced over
the last decade in the production of isotope-labeled samples, reconstitution of these
samples into the growing selection of NMR-compatible membrane-mimetic
systems, and the approaches used for the acquisition and application of structural
restraints from these complexes are reviewed.
Keywords Detergents
Global folds
Isotope labeling
NMR structure restraints
Protein expression
Contents
1 Introduction .................................................................................. 125
2 Production of Membrane Protein Samples for Solution NMR . . . ......................... 126
2.1 Factors Affecting Choice of Expression System ..................................... 126
2.2 Membrane Protein Expression in
E. Coli
............................................ 126
2.3 Cell-Free Expression Approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
