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Title: Structural prerequisites for G-protein activation by the neurotensin receptor

Abstract

We previously determined the structure of neurotensin receptor NTSR1 in an active-like conformation with six thermostabilizing mutations bound to the peptide agonist neurotensin. This receptor was unable to activate G proteins, indicating that the mutations restricted NTSR1 to relate agonist binding to G-protein activation. Here we analyse the effect of three of those mutations (E166A 3.49, L310A 6.37, F358A 7.42) and present two structures of NTSR1 able to catalyse nucleotide exchange at Gα. The presence of F358 7.42 causes the conserved W321 6.48 to adopt a side chain orientation parallel to the lipid bilayer sealing the collapsed Na+ ion pocket and linking the agonist with residues in the lower receptor part implicated in GPCR activation. In the intracellular receptor half, the bulkier L310 6.37 side chain dictates the position of R167 3.50 of the highly conserved D/ERY motif. These residues, together with the presence of E166 3.49 provide determinants for G-protein activation by NTSR1.

Authors:
 [1];  [1];  [1];  [1]
  1. National Inst. of Health (NIH), Rockville, MD (United States). Dept. of Health and Human Services
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS); SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1203746
Grant/Contract Number:  
AC02-06CH11357; AC02-76SF00515
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 6; Journal Issue: 07, 2015; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Krumm, Brian E., White, Jim F., Shah, Priyanka, and Grisshammer, Reinhard. Structural prerequisites for G-protein activation by the neurotensin receptor. United States: N. p., 2015. Web. doi:10.1038/ncomms8895.
Krumm, Brian E., White, Jim F., Shah, Priyanka, & Grisshammer, Reinhard. Structural prerequisites for G-protein activation by the neurotensin receptor. United States. doi:10.1038/ncomms8895.
Krumm, Brian E., White, Jim F., Shah, Priyanka, and Grisshammer, Reinhard. Fri . "Structural prerequisites for G-protein activation by the neurotensin receptor". United States. doi:10.1038/ncomms8895. https://www.osti.gov/servlets/purl/1203746.
@article{osti_1203746,
title = {Structural prerequisites for G-protein activation by the neurotensin receptor},
author = {Krumm, Brian E. and White, Jim F. and Shah, Priyanka and Grisshammer, Reinhard},
abstractNote = {We previously determined the structure of neurotensin receptor NTSR1 in an active-like conformation with six thermostabilizing mutations bound to the peptide agonist neurotensin. This receptor was unable to activate G proteins, indicating that the mutations restricted NTSR1 to relate agonist binding to G-protein activation. Here we analyse the effect of three of those mutations (E166A3.49, L310A6.37, F358A7.42) and present two structures of NTSR1 able to catalyse nucleotide exchange at Gα. The presence of F3587.42 causes the conserved W3216.48 to adopt a side chain orientation parallel to the lipid bilayer sealing the collapsed Na+ ion pocket and linking the agonist with residues in the lower receptor part implicated in GPCR activation. In the intracellular receptor half, the bulkier L3106.37 side chain dictates the position of R1673.50 of the highly conserved D/ERY motif. These residues, together with the presence of E1663.49 provide determinants for G-protein activation by NTSR1.},
doi = {10.1038/ncomms8895},
journal = {Nature Communications},
number = 07, 2015,
volume = 6,
place = {United States},
year = {2015},
month = {7}
}

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