Role of an Absolutely Conserved Tryptophan Pair in the Extracellular Domain of Cys-Loop Receptors

Nina Braun; Timothy Lynagh; Rilei Yu; Philip C Biggin; Stephan A Pless

doi:10.1021/acschemneuro.5b00298

Role of an Absolutely Conserved Tryptophan Pair in the Extracellular Domain of Cys-Loop Receptors

Nina Braun, Timothy Lynagh, Rilei Yu, Philip C Biggin, Stephan A Pless

5 Citations (Scopus)

Abstract

Cys-loop receptors mediate fast synaptic transmission in the nervous system, and their dysfunction is associated with a number of diseases. While some sequence variability is essential to ensure specific recognition of a chemically diverse set of ligands, other parts of the underlying amino acid sequences show a high degree of conservation, possibly to preserve the overall structural fold across the protein family. In this study, we focus on the only two absolutely conserved residues across the Cys-loop receptor family, two Trp side chains in the WXD motif of Loop D and in the WXPD motif of Loop A. Using a combination of conventional mutagenesis, unnatural amino acid incorporation, immunohistochemistry and MD simulations, we demonstrate the crucial contributions of these two Trp residues to receptor expression and function in two prototypical Cys-loop receptors, the anion-selective GlyR α1 and the cation-selective nAChR α7. Specifically, our results rule out possible electrostatic contributions of these Trp side chains and instead suggest that the overall size and shape of this aromatic pair is required in stabilizing the Cys-loop receptor extracellular domain.

Original language	English
Journal	A C S Chemical Neuroscience
Volume	7
Issue number	3
Pages (from-to)	339–348
Number of pages	10
ISSN	1948-7193
DOIs	https://doi.org/10.1021/acschemneuro.5b00298
Publication status	Published - 16 Mar 2016

Access to Document

10.1021/acschemneuro.5b00298

Cite this

@article{ae0e7dd2dff94913a256426026f046ae,

title = "Role of an Absolutely Conserved Tryptophan Pair in the Extracellular Domain of Cys-Loop Receptors",

abstract = "Cys-loop receptors mediate fast synaptic transmission in the nervous system, and their dysfunction is associated with a number of diseases. While some sequence variability is essential to ensure specific recognition of a chemically diverse set of ligands, other parts of the underlying amino acid sequences show a high degree of conservation, possibly to preserve the overall structural fold across the protein family. In this study, we focus on the only two absolutely conserved residues across the Cys-loop receptor family, two Trp side chains in the WXD motif of Loop D and in the WXPD motif of Loop A. Using a combination of conventional mutagenesis, unnatural amino acid incorporation, immunohistochemistry and MD simulations, we demonstrate the crucial contributions of these two Trp residues to receptor expression and function in two prototypical Cys-loop receptors, the anion-selective GlyR α1 and the cation-selective nAChR α7. Specifically, our results rule out possible electrostatic contributions of these Trp side chains and instead suggest that the overall size and shape of this aromatic pair is required in stabilizing the Cys-loop receptor extracellular domain.",

author = "Nina Braun and Timothy Lynagh and Rilei Yu and Biggin, {Philip C} and Pless, {Stephan A}",

year = "2016",

month = mar,

day = "16",

doi = "10.1021/acschemneuro.5b00298",

language = "English",

volume = "7",

pages = " 339–348",

journal = "ACS Chemical Neuroscience",

issn = "1948-7193",

publisher = "American Chemical Society",

number = "3",

}

TY - JOUR

T1 - Role of an Absolutely Conserved Tryptophan Pair in the Extracellular Domain of Cys-Loop Receptors

AU - Braun, Nina

AU - Lynagh, Timothy

AU - Yu, Rilei

AU - Biggin, Philip C

AU - Pless, Stephan A

PY - 2016/3/16

Y1 - 2016/3/16

N2 - Cys-loop receptors mediate fast synaptic transmission in the nervous system, and their dysfunction is associated with a number of diseases. While some sequence variability is essential to ensure specific recognition of a chemically diverse set of ligands, other parts of the underlying amino acid sequences show a high degree of conservation, possibly to preserve the overall structural fold across the protein family. In this study, we focus on the only two absolutely conserved residues across the Cys-loop receptor family, two Trp side chains in the WXD motif of Loop D and in the WXPD motif of Loop A. Using a combination of conventional mutagenesis, unnatural amino acid incorporation, immunohistochemistry and MD simulations, we demonstrate the crucial contributions of these two Trp residues to receptor expression and function in two prototypical Cys-loop receptors, the anion-selective GlyR α1 and the cation-selective nAChR α7. Specifically, our results rule out possible electrostatic contributions of these Trp side chains and instead suggest that the overall size and shape of this aromatic pair is required in stabilizing the Cys-loop receptor extracellular domain.

AB - Cys-loop receptors mediate fast synaptic transmission in the nervous system, and their dysfunction is associated with a number of diseases. While some sequence variability is essential to ensure specific recognition of a chemically diverse set of ligands, other parts of the underlying amino acid sequences show a high degree of conservation, possibly to preserve the overall structural fold across the protein family. In this study, we focus on the only two absolutely conserved residues across the Cys-loop receptor family, two Trp side chains in the WXD motif of Loop D and in the WXPD motif of Loop A. Using a combination of conventional mutagenesis, unnatural amino acid incorporation, immunohistochemistry and MD simulations, we demonstrate the crucial contributions of these two Trp residues to receptor expression and function in two prototypical Cys-loop receptors, the anion-selective GlyR α1 and the cation-selective nAChR α7. Specifically, our results rule out possible electrostatic contributions of these Trp side chains and instead suggest that the overall size and shape of this aromatic pair is required in stabilizing the Cys-loop receptor extracellular domain.

U2 - 10.1021/acschemneuro.5b00298

DO - 10.1021/acschemneuro.5b00298

M3 - Journal article

C2 - 26764897

SN - 1948-7193

VL - 7

SP - 339

EP - 348

JO - ACS Chemical Neuroscience

JF - ACS Chemical Neuroscience

IS - 3

ER -

Role of an Absolutely Conserved Tryptophan Pair in the Extracellular Domain of Cys-Loop Receptors

Abstract

Access to Document

Fingerprint

Cite this