Tyrosine 288 in the extracellular domain of the human P2X7 receptor is critical for receptor function revealed by structural modelling and site‐directed mutagenesis

2021 ◽  
Author(s):  
Emily A. Caseley ◽  
Stephen P. Muench ◽  
Lin‐Hua Jiang
Keyword(s):  
P2x7 Receptor ◽  
Extracellular Domain ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Structural Modelling ◽  
Receptor Function

Role of Cysteine Residues in the N-Terminal Extracellular Domain of the Human VIP 1 Receptor for Ligand Binding A Site-directed Mutagenesis Studya

2006 ◽  
Vol 805 (1) ◽  
pp. 585-589 ◽  
Author(s):  
PASCALE GAUDIN ◽  
ALAIN COUVINEAU ◽  
JEAN-JOSÉ MAORET ◽  
CHRISTIANE ROUYER-FESSARD ◽  
MARC LABURTHE
Keyword(s):  
Ligand Binding ◽  
Extracellular Domain ◽  
Site Directed Mutagenesis ◽  
Cysteine Residues ◽  
Directed Mutagenesis ◽  
A Site

scholarly journals Structural basis for the functional properties of the P2X7 receptor for extracellular ATP

2021 ◽  
Author(s):  
Lin-Hua Jiang ◽  
Emily A. Caseley ◽  
Steve P. Muench ◽  
Sébastien Roger
Keyword(s):  
Functional Properties ◽  
P2x7 Receptor ◽  
Site Directed Mutagenesis ◽  
P2x Receptor ◽  
Structural Basis ◽  
Structure Characteristic ◽  
Purinergic Signalling ◽  
Directed Mutagenesis ◽  
Important Mediator ◽  
Receptor Family

AbstractThe P2X7 receptor, originally known as the P2Z receptor due to its distinctive functional properties, has a structure characteristic of the ATP-gated ion channel P2X receptor family. The P2X7 receptor is an important mediator of ATP-induced purinergic signalling and is involved the pathogenesis of numerous conditions as well as in the regulation of diverse physiological functions. Functional characterisations, in conjunction with site-directed mutagenesis, molecular modelling, and, recently, structural determination, have provided significant insights into the structure–function relationships of the P2X7 receptor. This review discusses the current understanding of the structural basis for the functional properties of the P2X7 receptor.

Ouabain-resistant mutants of the rat Na,K-ATPase alpha 2 isoform identified by using an episomal expression vector

1990 ◽  
Vol 10 (4) ◽  
pp. 1367-1372
Author(s):  
V Canfield ◽  
J R Emanuel ◽  
N Spickofsky ◽  
R Levenson ◽  
R F Margolskee
Keyword(s):  
Partial Resistance ◽  
Expression Vector ◽  
Extracellular Domain ◽  
Site Directed Mutagenesis ◽  
Transformed Cells ◽  
Directed Mutagenesis ◽  
Resistance Level ◽  
Resistant Mutants ◽  
Double Substitution ◽  
Ouabain Sensitivity

Site-directed mutagenesis was used to identify residues responsible for the greater than 1,000-fold difference in ouabain sensitivity between the rat Na,K-ATPase alpha 1 and alpha 2 isoforms. A series of mutagenized cDNAs was constructed that replaced residues of the rat alpha 2 subunit with the corresponding residues from the rat alpha 1 subunit. These cDNAs were cloned into a mammalian episomal expression vector (EBOpLPP) and expressed in ouabain-sensitive primate cells. Either of two single substitutions introduced into the rat alpha 2 subunit cDNA (Leu-111----Arg or Asn-122----Asp) conferred partial resistance (approximately 10 microM ouabain) upon transformed cells. This resistance was intermediate between the levels conferred by the rat alpha 1 cDNA (approximately 500 microM ouabain) and the rat alpha 2 cDNA (approximately 0.2 microM ouabain). A double substitution of the rat alpha 2 cDNA (Leu-111----Arg and Asn-122----Asp) conferred a resistance level equivalent to that obtained with rat alpha 1. These results demonstrate that the residues responsible for isoform-specific differences in ouabain sensitivity are located at the end of the H1-H2 extracellular domain. The combination of site-directed mutagenesis and episomal expression provides a useful system for the selection and analysis of mutants.

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