Rhodopsin recognition by mutant G(s)α containing C-terminal residues of transducin

Michael Natochin, Khakim G. Muradov, Randall Lee McEntaffer, Nikolai O. Artemyev

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The C-terminal regions of the heterotrimeric G protein α-subunits play key roles in selective activation of G proteins by their cognate receptors. In this study, mutant G(s)α proteins with substitutions by C-terminal residues of transducin (G(t)α) were analyzed for their interaction with light-activated rhodopsin (R*) to delineate the critical determinants of the G(t)α/R* coupling. In contrast to G(s)α, a chimeric G(s)α/G(t)α protein containing only 11 C-terminal residues from transducin was capable of binding to and being potently activated by R(*). Our results suggest that Cys347 and Gly348 are absolutely essential, whereas Asp346 is more modestly involved in the G(t) activation by R(*). In addition, the analysis of the intrinsic nucleotide exchange in mutant G(s)α indicated an interaction between the C terminus and the switch II region in G(t)α-GDP. Mutant G(s)α containing the G(t)α C terminus and substitutions of Asn239 and Asp240 (switch II) by the corresponding G(t)α residues, Glu212 and Gly213, displayed significant reductions in spontaneous guanosine 5'-O-(3- thiotriphosphate)-binding rates to the levels approaching those in G(t)α. Communication between the C terminus and switch II of G(t)α does not appear essential for the activational coupling between G(t) and R*, but may represent one of the mechanisms by which Gα subunits control intrinsic nucleotide exchange.

Original languageEnglish (US)
Pages (from-to)2669-2675
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number4
DOIs
StatePublished - Jan 28 2000

Fingerprint

Transducin
Rhodopsin
Nucleotides
Switches
Guanosine 5'-O-(3-Thiotriphosphate)
Heterotrimeric GTP-Binding Proteins
Protein Subunits
Substitution reactions
Chemical activation
GTP-Binding Proteins
Proteins
Light
Communication

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Natochin, Michael ; Muradov, Khakim G. ; McEntaffer, Randall Lee ; Artemyev, Nikolai O. / Rhodopsin recognition by mutant G(s)α containing C-terminal residues of transducin. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 4. pp. 2669-2675.
@article{afae46a305d84a76877799515d5d4e17,
title = "Rhodopsin recognition by mutant G(s)α containing C-terminal residues of transducin",
abstract = "The C-terminal regions of the heterotrimeric G protein α-subunits play key roles in selective activation of G proteins by their cognate receptors. In this study, mutant G(s)α proteins with substitutions by C-terminal residues of transducin (G(t)α) were analyzed for their interaction with light-activated rhodopsin (R*) to delineate the critical determinants of the G(t)α/R* coupling. In contrast to G(s)α, a chimeric G(s)α/G(t)α protein containing only 11 C-terminal residues from transducin was capable of binding to and being potently activated by R(*). Our results suggest that Cys347 and Gly348 are absolutely essential, whereas Asp346 is more modestly involved in the G(t) activation by R(*). In addition, the analysis of the intrinsic nucleotide exchange in mutant G(s)α indicated an interaction between the C terminus and the switch II region in G(t)α-GDP. Mutant G(s)α containing the G(t)α C terminus and substitutions of Asn239 and Asp240 (switch II) by the corresponding G(t)α residues, Glu212 and Gly213, displayed significant reductions in spontaneous guanosine 5'-O-(3- thiotriphosphate)-binding rates to the levels approaching those in G(t)α. Communication between the C terminus and switch II of G(t)α does not appear essential for the activational coupling between G(t) and R*, but may represent one of the mechanisms by which Gα subunits control intrinsic nucleotide exchange.",
author = "Michael Natochin and Muradov, {Khakim G.} and McEntaffer, {Randall Lee} and Artemyev, {Nikolai O.}",
year = "2000",
month = "1",
day = "28",
doi = "10.1074/jbc.275.4.2669",
language = "English (US)",
volume = "275",
pages = "2669--2675",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "4",

}

Rhodopsin recognition by mutant G(s)α containing C-terminal residues of transducin. / Natochin, Michael; Muradov, Khakim G.; McEntaffer, Randall Lee; Artemyev, Nikolai O.

In: Journal of Biological Chemistry, Vol. 275, No. 4, 28.01.2000, p. 2669-2675.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Rhodopsin recognition by mutant G(s)α containing C-terminal residues of transducin

AU - Natochin, Michael

AU - Muradov, Khakim G.

AU - McEntaffer, Randall Lee

AU - Artemyev, Nikolai O.

PY - 2000/1/28

Y1 - 2000/1/28

N2 - The C-terminal regions of the heterotrimeric G protein α-subunits play key roles in selective activation of G proteins by their cognate receptors. In this study, mutant G(s)α proteins with substitutions by C-terminal residues of transducin (G(t)α) were analyzed for their interaction with light-activated rhodopsin (R*) to delineate the critical determinants of the G(t)α/R* coupling. In contrast to G(s)α, a chimeric G(s)α/G(t)α protein containing only 11 C-terminal residues from transducin was capable of binding to and being potently activated by R(*). Our results suggest that Cys347 and Gly348 are absolutely essential, whereas Asp346 is more modestly involved in the G(t) activation by R(*). In addition, the analysis of the intrinsic nucleotide exchange in mutant G(s)α indicated an interaction between the C terminus and the switch II region in G(t)α-GDP. Mutant G(s)α containing the G(t)α C terminus and substitutions of Asn239 and Asp240 (switch II) by the corresponding G(t)α residues, Glu212 and Gly213, displayed significant reductions in spontaneous guanosine 5'-O-(3- thiotriphosphate)-binding rates to the levels approaching those in G(t)α. Communication between the C terminus and switch II of G(t)α does not appear essential for the activational coupling between G(t) and R*, but may represent one of the mechanisms by which Gα subunits control intrinsic nucleotide exchange.

AB - The C-terminal regions of the heterotrimeric G protein α-subunits play key roles in selective activation of G proteins by their cognate receptors. In this study, mutant G(s)α proteins with substitutions by C-terminal residues of transducin (G(t)α) were analyzed for their interaction with light-activated rhodopsin (R*) to delineate the critical determinants of the G(t)α/R* coupling. In contrast to G(s)α, a chimeric G(s)α/G(t)α protein containing only 11 C-terminal residues from transducin was capable of binding to and being potently activated by R(*). Our results suggest that Cys347 and Gly348 are absolutely essential, whereas Asp346 is more modestly involved in the G(t) activation by R(*). In addition, the analysis of the intrinsic nucleotide exchange in mutant G(s)α indicated an interaction between the C terminus and the switch II region in G(t)α-GDP. Mutant G(s)α containing the G(t)α C terminus and substitutions of Asn239 and Asp240 (switch II) by the corresponding G(t)α residues, Glu212 and Gly213, displayed significant reductions in spontaneous guanosine 5'-O-(3- thiotriphosphate)-binding rates to the levels approaching those in G(t)α. Communication between the C terminus and switch II of G(t)α does not appear essential for the activational coupling between G(t) and R*, but may represent one of the mechanisms by which Gα subunits control intrinsic nucleotide exchange.

UR - http://www.scopus.com/inward/record.url?scp=0034723294&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034723294&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.4.2669

DO - 10.1074/jbc.275.4.2669

M3 - Article

C2 - 10644728

AN - SCOPUS:0034723294

VL - 275

SP - 2669

EP - 2675

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 4

ER -