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Approaches towards Structures of Y Receptors, Examples of Human G-Protein Coupled Receptors, by Solution NMR


Walser, R; Zerbe, O (2012). Approaches towards Structures of Y Receptors, Examples of Human G-Protein Coupled Receptors, by Solution NMR. CHIMIA International Journal for Chemistry, 66(10):781-786.

Abstract

Despite recent advances no solution structure for a true G-protein coupled receptor (GPCR) is available today due to biochemical and spectroscopic problems. Herein we review our attempts to obtain assignments of GPCRs based on fragments comprising 2-3 transmembrane helices. The fragments are expressed in a heterologous system, and studied in detergent micelles using solution NMR spectroscopy. We report on the status of assignments of fragments from the Y4 receptor, a human GPCR. Assignments for the majority of the backbone resonances are available as well as sidechain assignments for the first two TM helices. Residues of TM4 are largely invisible. We review technical issues in preparing these samples and in the data analysis. In addition we developed an approach in which we have grafted the extracellular loops of the Y1 receptor onto a beta-barrel scaffold derived from the E. coli outer membrane protein OmpA. We could demonstrate that all loops can be successfully transferred, and that the resulting protein can be successfully refolded. The system is capable of recognizing the ligands from the neuropeptide Y family.

Abstract

Despite recent advances no solution structure for a true G-protein coupled receptor (GPCR) is available today due to biochemical and spectroscopic problems. Herein we review our attempts to obtain assignments of GPCRs based on fragments comprising 2-3 transmembrane helices. The fragments are expressed in a heterologous system, and studied in detergent micelles using solution NMR spectroscopy. We report on the status of assignments of fragments from the Y4 receptor, a human GPCR. Assignments for the majority of the backbone resonances are available as well as sidechain assignments for the first two TM helices. Residues of TM4 are largely invisible. We review technical issues in preparing these samples and in the data analysis. In addition we developed an approach in which we have grafted the extracellular loops of the Y1 receptor onto a beta-barrel scaffold derived from the E. coli outer membrane protein OmpA. We could demonstrate that all loops can be successfully transferred, and that the resulting protein can be successfully refolded. The system is capable of recognizing the ligands from the neuropeptide Y family.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Language:English
Date:2012
Deposited On:05 Nov 2012 16:57
Last Modified:07 Dec 2017 16:01
Publisher:Swiss Chemical Society
ISSN:0009-4293
Additional Information:Copyright ©Swiss Chemical Society: CHIMIA, 2012, 66/10 S. 781-786
Publisher DOI:https://doi.org/10.2533/chimia.2012.781

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