Evidence map›Paper›PMID 27002154›Full record

ArticleThe Journal of biological chemistry2016

Intrinsic Pleckstrin Homology (PH) Domain Motion in Phospholipase C-β Exposes a Gβγ Protein Binding Site.

Ganesh Kadamur, Elliott M Ross

Open access · hybridAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in The Journal of biological chemistry, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.

0numbers the graph read from it
0cells of the map it votes in
12citing papers in PubMed
0.7field-weighted citation impact, top 26% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

12 citing papers in PubMed, 21 citations in OpenAlex.

  1. Article
  2. Article
  3. Article
  4. Proceedings of the National Academy of Sciences of the United States of America · 2023
    Article
  5. Molecular regulation of PLCβ signaling.Methods in enzymology · 2023
    Review
  6. Article
  7. Review
  8. Article
  9. Activation of Phospholipase C β by Gβγ and GαStructure (London, England : 1993) · 2020
    Article
  10. Article
  11. The binding of activated GαThe Journal of biological chemistry · 2017
    Article
  12. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

2 authors at 2 institutions in 1 country.

Ganesh KadamurFrom the Department of Pharmacology, Molecular Biophysics Graduate Program, and Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Elliott M RossFrom the Department of Pharmacology, Molecular Biophysics Graduate Program, and Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390 Elliott.Ross@UTSouthwestern.edu.ORCID http://orcid.org/0000-0002-0252-8197
Center for Systems Biology · USSouthwestern Medical Center · US

Funding

G Protein-Coupled Receptors-Structure and RegulationR01GM030355 · NIGMS · UT SOUTHWESTERN MEDICAL CENTER · PI ROSS, ELLIOTT M · 1985 to 2015
$5.0M
NIGMS NIH HHS R01 GM030355
6 · The paper itself

Abstract

Mammalian phospholipase C-β (PLC-β) isoforms are stimulated by heterotrimeric G protein subunits and members of the Rho GTPase family of small G proteins. Although recent structural studies showed how Gαq and Rac1 bind PLC-β, there is a lack of consensus regarding the Gβγ binding site in PLC-β. Using FRET between cerulean fluorescent protein-labeled Gβγ and the Alexa Fluor 594-labeled PLC-β pleckstrin homology (PH) domain, we demonstrate that the PH domain is the minimal Gβγ binding region in PLC-β3. We show that the isolated PH domain can compete with full-length PLC-β3 for binding Gβγ but not Gαq, Using sequence conservation, structural analyses, and mutagenesis, we identify a hydrophobic face of the PLC-β PH domain as the Gβγ binding interface. This PH domain surface is not solvent-exposed in crystal structures of PLC-β, necessitating conformational rearrangement to allow Gβγ binding. Blocking PH domain motion in PLC-β by cross-linking it to the EF hand domain inhibits stimulation by Gβγ without altering basal activity or Gαq response. The fraction of PLC-β cross-linked is proportional to the fractional loss of Gβγ response. Cross-linked PLC-β does not bind Gβγ in a FRET-based Gβγ-PLC-β binding assay. We propose that unliganded PLC-β exists in equilibrium between a closed conformation observed in crystal structures and an open conformation where the PH domain moves away from the EF hands. Therefore, intrinsic movement of the PH domain in PLC-β modulates Gβγ access to its binding site.

Indexed as

Binding SitesBlood ProteinsFluorescence Resonance Energy TransferGTP-Binding Protein beta SubunitsGTP-Binding Protein gamma SubunitsHumansModels, MolecularPhospholipase C betaPhosphoproteinsProtein BindingProtein Interaction Domains and MotifsProtein Structure, Tertiaryrac1 GTP-Binding ProteinRecombinant ProteinsStructural Homology, ProteinBlood ProteinsGTP-Binding Protein beta SubunitsGTP-Binding Protein gamma SubunitsPhospholipase C betaPhosphoproteinsplatelet protein P47PLCB3 protein, humanrac1 GTP-Binding ProteinRAC1 protein, humanRecombinant Proteinscell signalingGβγheterotrimeric G proteinPH domainphosphatidylinositol signalingphospholipase Cprotein conformation

Identifiers

PMID27002154
PMCPMC4900283
OpenAlexW2408445002

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.