The guanine nucleotide exchange factor Ric-8a induces domain separation and Ras domain plasticity in Gαi1

Ned Van Eps, Celestine J. Thomas, Wayne L. Hubbell, Stephen R. Sprang

Research output: Contribution to journalArticlepeer-review

Abstract

Heterotrimeric G proteins are activated by exchange of GDP for GTP at the G protein alpha subunit (Gα), most notably by G pro-tein-coupled transmembrane receptors. Ric-8A is a soluble cytoplasmic protein essential for embryonic development that acts as both a guanine nucleotide exchange factor (GEF) and a chaperone for Gα subunits of the i, q, and 12/13 classes. Previous studies demonstrated that Ric-8A stabilizes a dynamically disordered state of nucleotide-free Gα as the catalytic intermediate for nucleotide exchange, but no information was obtained on the structures involved or the magnitude of the structural fluctuations. In the present study, site-directed spin labeling (SDSL) together with double electron-electron resonance (DEER) spectroscopy is used to provide global distance constraints that identify discrete members of a conformational ensemble in the Gαi1:Ric-8A complex and the magnitude of structural differences between them. In the complex, the helical and Ras-like nucleotide-binding domains of Gαi1 pivot apart to occupy multiple resolved states with displacements as large as 25 Å. The domain displacement appears to be distinct from that observed in Gαs upon binding of Gs to the β2 adrenergic receptor. Moreover, the Ras-like domain exhibits structural plasticity within and around the nucleotide-binding cavity, and the switch I and switch II regions, which are known to adopt different conformations in the GDP- and GTP-bound states of Gα, undergo structural rearrangements. Collectively, the data show that Ric-8A induces a conformationally heterogeneous state of Gαi and provide insight into the mechanism of action of a nonreceptor Gα GEF.

Original languageEnglish
Pages (from-to)1404-1409
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number5
DOIs
StatePublished - Feb 3 2015

Keywords

  • Double electron electron resonance spectroscopy
  • G protein
  • Guanine nucleotide exchange factor
  • Protein dynamics
  • Tertiary structure

Fingerprint

Dive into the research topics of 'The guanine nucleotide exchange factor Ric-8a induces domain separation and Ras domain plasticity in Gαi1'. Together they form a unique fingerprint.

Cite this