Format

Send to

Choose Destination
J Biol Chem. 2014 Jun 20;289(25):17791-801. doi: 10.1074/jbc.M114.576827. Epub 2014 May 7.

A chemical biology approach demonstrates G protein βγ subunits are sufficient to mediate directional neutrophil chemotaxis.

Author information

1
From the Departments of Biochemistry and Biophysics and.
2
Pharmacology and Physiology, University of Rochester, Rochester, New York 14642.
3
From the Departments of Biochemistry and Biophysics and Pharmacology and Physiology, University of Rochester, Rochester, New York 14642 alan_smrcka@urmc.rochester.edu.

Abstract

Our laboratory has identified a number of small molecules that bind to G protein βγ subunits (Gβγ) by competing for peptide binding to the Gβγ "hot spot." M119/Gallein were identified as inhibitors of Gβγ subunit signaling. Here we examine the activity of another molecule identified in this screen, 12155, which we show that in contrast to M119/Gallein had no effect on Gβγ-mediated phospholipase C or phosphoinositide 3-kinase (PI3K) γ activation in vitro. Also in direct contrast to M119/Gallein, 12155 caused receptor-independent Ca(2+) release, and activated other downstream targets of Gβγ including extracellular signal regulated kinase (ERK), protein kinase B (Akt) in HL60 cells differentiated to neutrophils. We show that 12155 releases Gβγ in vitro from Gαi1β1γ2 heterotrimers by causing its dissociation from GαGDP without inducing nucleotide exchange in the Gα subunit. We used this novel probe to examine the hypothesis that Gβγ release is sufficient to direct chemotaxis of neutrophils in the absence of receptor or G protein α subunit activation. 12155 directed chemotaxis of HL60 cells and primary neutrophils in a transwell migration assay with responses similar to those seen for the natural chemotactic peptide n-formyl-Met-Leu-Phe. These data indicate that release of free Gβγ is sufficient to drive directional chemotaxis in a G protein-coupled receptor signaling-independent manner.

KEYWORDS:

Cell Signaling; Chemical Biology; Chemotaxis; G Protein; G Protein-coupled Receptor (GPCR)

PMID:
24808183
PMCID:
PMC4067212
DOI:
10.1074/jbc.M114.576827
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center