Expression of Double Transmembrane Domain GPCR Fragments for Biophysical Analysis
There are thousands of protein structures in the databases that have been determined using X-ray crystallography and nuclear magnetic resonance spectroscopy. Despite the prevalence of integral membrane proteins, or IMPs (20-30% of the human genome encodes IMPs) few structures of these proteins have been determined, and only one G protein-coupled receptors (GPCRs) structure, that of rhodopsin, has been solved by X-ray crystallography . Based on biochemical and biophysical analyses, GPCRs have been shown to consist of an N-terminal extracellular region, seven transmembrane (TM) helices separated by intra- and extra-cellular loops, and a C-terminal cytoplasmic tail. One of the factors that limits the number of high resolution structures is the low level of expression for a full-length GPCR. A few groups have started to study fragments of GPCRs to help to elucidate their structures [2, 3].
We have recently published high resolution structures of a 73-residue peptide...
KeywordsMolar Ellipticity High Resolution Structure Biophysical Analysis Organic Aqueous Medium CNBr Cleavage
This work was supported by grants NIH GM-22086 and NIH GM-22087 to FN and JMB.
- 5.Neumoin, A., Arshava, B., Becker, J., Zerbe, O., and Naider, F. Biophys J (2007).Google Scholar