Measurement of GTPγS binding to specific G proteins in membranes using G-protein antibodies

FEBS Letters ◽  
1992 ◽  
Vol 305 (2) ◽  
pp. 125-128 ◽  
Author(s):  
Takashi Okamoto ◽  
Tsuneya Ikezu ◽  
Yoshitake Murayama ◽  
Etsuro Ogata ◽  
Ikuo Nishimoto
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Gtpγs Binding

A study of M1 receptor coupling to inhibitory G proteins in CHO cells via GTPγS binding determined with anti-G protein antibodies

Life Sciences ◽  
1999 ◽  
Vol 64 (6-7) ◽  
pp. 569 ◽  
Author(s):  
N.W. DeLapp ◽  
D. McClure ◽  
J. McKinzie ◽  
A. Silbernagel ◽  
B.D. Sawyer ◽  
...  
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Cho Cells ◽  
Receptor Coupling ◽  
M1 Receptor ◽  
Gtpγs Binding

scholarly journals Every Detail Matters. That Is, How the Interaction between Gα Proteins and Membrane Affects Their Function

Membranes ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 222
Author(s):  
Agnieszka Polit ◽  
Paweł Mystek ◽  
Ewa Błasiak
Keyword(s):  
Signal Transduction ◽  
G Protein ◽  
G Proteins ◽  
Lipid Membranes ◽  
Signaling Proteins ◽  
Heterotrimeric G Proteins ◽  
Membrane Attachment ◽  
The Individual ◽  
Multicellular Organisms ◽  
G Protein Coupled

In highly organized multicellular organisms such as humans, the functions of an individual cell are dependent on signal transduction through G protein-coupled receptors (GPCRs) and subsequently heterotrimeric G proteins. As most of the elements belonging to the signal transduction system are bound to lipid membranes, researchers are showing increasing interest in studying the accompanying protein–lipid interactions, which have been demonstrated to not only provide the environment but also regulate proper and efficient signal transduction. The mode of interaction between the cell membrane and G proteins is well known. Despite this, the recognition mechanisms at the molecular level and how the individual G protein-membrane attachment signals are interrelated in the process of the complex control of membrane targeting of G proteins remain unelucidated. This review focuses on the mechanisms by which mammalian Gα subunits of G proteins interact with lipids and the factors responsible for the specificity of membrane association. We summarize recent data on how these signaling proteins are precisely targeted to a specific site in the membrane region by introducing well-defined modifications as well as through the presence of polybasic regions within these proteins and interactions with other components of the heterocomplex.

scholarly journals Production, processing and partial purification of functional G protein βγ subunits in baculovirus-infected insect cells

1992 ◽  
Vol 286 (3) ◽  
pp. 677-680 ◽  
Author(s):  
J D Robishaw ◽  
V K Kalman ◽  
K L Proulx
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Partial Purification ◽  
Baculovirus Expression ◽  
Gamma Subunits ◽  
Infected Insect ◽  
Beta 2

As a result of the inability to resolve the heterogeneous mixture of G protein beta gamma subunits present in tissues, it has not been possible to compare different beta gamma subunits of the G proteins in terms of their proposed roles in receptor-effector coupling. This study was undertaken to establish the utility of the baculovirus expression system in producing homogeneous beta gamma subunits of defined composition for the comparative analysis of these subunits in reconstitution systems. In this study we report the expression, and appropriate post-translational processing, of recombinant beta 2, gamma 2 and gamma 3 subunits. In addition, we show that the recombinant beta gamma subunits can be readily purified, and can functionally interact with the alpha subunits of the G proteins.

scholarly journals Signaling from G Protein-coupled Receptors to ERK5/Big MAPK 1 Involves Gαqand Gα12/13Families of Heterotrimeric G Proteins

2000 ◽  
Vol 275 (28) ◽  
pp. 21730-21736 ◽  
Author(s):  
Shigetomo Fukuhara ◽  
Maria Julia Marinissen ◽  
Mario Chiariello ◽  
J. Silvio Gutkind
Keyword(s):  
G Protein ◽  
G Proteins ◽  
G Protein Coupled Receptors ◽  
Heterotrimeric G Proteins ◽  
G Protein Coupled

scholarly journals G Protein–Coupled Receptor-Type G Proteins Are Required for Light-Dependent Seedling Growth and Fertility in Arabidopsis

2012 ◽  
Vol 24 (9) ◽  
pp. 3649-3668 ◽  
Author(s):  
Felix W. Jaffé ◽  
Gian-Enrico C. Freschet ◽  
Billy M. Valdes ◽  
John Runions ◽  
Matthew J. Terry ◽  
...  
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Seedling Growth ◽  
Receptor Type ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

scholarly journals Modulation of Ins(2,4,5)P3-stimulated Ca2+ mobilization by Ins(1,3,4,5)P4: enhancement by activated G-proteins, and evidence for the involvement of a GAP1 protein, a putative Ins(1,3,4,5)P4 receptor

1998 ◽  
Vol 331 (3) ◽  
pp. 947-952 ◽  
Author(s):  
Jefferson W. LOOMIS-HUSSELBEE ◽  
Christopher D. WALKER ◽  
Joanna R. BOTTOMLEY ◽  
Peter J. CULLEN ◽  
Robin F. IRVINE ◽  
...  
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Protein A ◽  
Related Protein ◽  
L1210 Cells ◽  
Putative Receptor ◽  
Specific Enhancement ◽  
Insp3 Receptor

We have previously shown that addition of Ins(1,3,4,5)P4 to permeabilized L1210 cells increases the amount of Ca2+ mobilized by a submaximal concentration of Ins(2,4,5)P3, and we suggested that, in doing this, Ins(1,3,4,5)P4 is not working via an InsP3 receptor but indirectly via an InsP4 receptor [Loomis-Husselbee, Cullen, Dreikhausen, Irvine and Dawson (1996) Biochem. J. 314, 811–816]. Here we have investigated whether this effect might be mediated by GAP1IP4BP, recently identified as a putative receptor for Ins(1,3,4,5)P4. GAP1IP4BP is a protein that interacts with one or more monomeric G-proteins, so we sought evidence for involvement of monomeric G-proteins in the effects of Ins(1,3,4,5)P4 in permeabilized L1210 cells. Guanosine 5´-[γ-thio]triphosphate (GTP[S]) enhanced the effect of Ins(1,3,4,5)P4 on Ins(2,4,5)P3-stimulated Ca2+ mobilization, but had no effect on the action of Ins(2,4,5)P3 alone. A specific enhancement of only the action of Ins(1,3,4,5)P4 was also seen with GTP[S]-loaded R-Ras or Rap1a (two G-proteins known to interact with GAP1IP4BP), whereas H-Ras was inactive at similar concentrations. Guanosine 5´-[β-thio]diphosphate (GDP[S]) did not alter the action of either Ins(2,4,5)P3 or Ins(1,3,4,5)P4. Finally, the addition of exogenous GAP1IP4BP, purified from platelets, markedly enhanced the effect of Ins(1,3,4,5)P4, and again, the amount of Ca2+ mobilized by Ins(2,4,5)P3 alone was unaltered. We conclude that the increase in Ins(2,4,5)P3-stimulated Ca2+ mobilization by Ins(1,3,4,5)P4 may be mediated by GAP1IP4BP or a closely related protein (such as GAP1m), and if so, the action of the GAP1 is not solely to regulate GTP loading of a G-protein, but rather it acts with a G-protein to cause its effect.

Sign in / Sign up

Export Citation Format

Share Document