title: Cell-free protein synthesis as a tool to study RXFP3-relaxin-3 protein interactions.
abstract: With the discovery of the relaxin family peptide receptors there is interest in obtaining a clearer understanding of the structure of these proteins and the molecular mechanism of receptor-ligand interaction. As G-protein coupled receptors, obtaining milligram quantities for structural investigations is hampered by the inherent instability of these integral membrane proteins. In the current context, understanding of GPCR structural biology has increased dramatically with crystal structures of several inactive and now active forms solved. In addition, the first nuclear magnetic resonance structure of a GPCR was obtained which is of crucial importance to studying these receptors in a more "biologically relevant" setting. However despite this expansion in the field, most structures have been solved on modified systems so as to increase stability and are not necessarily representative of the native receptors. In relation to the relaxin family peptide receptors, we chose to investigate relaxin-family peptide receptor-3 expressed by cell-free protein synthesis. In contrast to in-vivo expression, cell-free was capable of producing large amounts of native receptor which makes it amenable to demanding structural studies.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Shilling
authors.forename: Ross A D
authors.initials: RA
authors.lastname: Bathgate
authors.forename: Paul R
authors.initials: PR
authors.lastname: Gooley
mesh_terms: D002474:Cell-Free System; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D054730:Protein Interaction Domains and Motifs; D055550:Protein Stability; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D012065:Relaxin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: C480238:RLN3 protein, human; C410092:RXFP3 protein, human; D043562:Receptors, G-Protein-Coupled; D012065:Relaxin