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TitleStructural underpinnings of Ric8A function as a G-protein α-subunit chaperone and guanine-nucleotide exchange factor.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 3084, Year 2019
Publish dateJul 12, 2019
AuthorsDhiraj Srivastava / Lokesh Gakhar / Nikolai O Artemyev /
PubMed AbstractResistance to inhibitors of cholinesterase 8A (Ric8A) is an essential regulator of G protein α-subunits (Gα), acting as a guanine nucleotide exchange factor and a chaperone. We report two crystal ...Resistance to inhibitors of cholinesterase 8A (Ric8A) is an essential regulator of G protein α-subunits (Gα), acting as a guanine nucleotide exchange factor and a chaperone. We report two crystal structures of Ric8A, one in the apo form and the other in complex with a tagged C-terminal fragment of Gα. These structures reveal two principal domains of Ric8A: an armadillo-fold core and a flexible C-terminal tail. Additionally, they show that the Gα C-terminus binds to a highly-conserved patch on the concave surface of the Ric8A armadillo-domain, with selectivity determinants residing in the Gα sequence. Biochemical analysis shows that the Ric8A C-terminal tail is critical for its stability and function. A model of the Ric8A/Gα complex derived from crosslinking mass spectrometry and molecular dynamics simulations suggests that the Ric8A C-terminal tail helps organize the GTP-binding site of Gα. This study lays the groundwork for understanding Ric8A function at the molecular level.
External linksNat Commun / PubMed:31300652 / PubMed Central
MethodsSAS (X-ray synchrotron) / X-ray diffraction
Resolution1.75 - 3.9 Å
Structure data

SASDF65:
Resistance to inhibitors of cholinesterase 8 homolog A (Ric8a) amino acids 1-492
Method: SAXS/SANS

SASDF75:
Resistance to inhibitors of cholinesterase 8 homolog A (Ric8a) amino acids 1-452
Method: SAXS/SANS

PDB-6n84:
MBP-fusion protein of transducin-alpha residues 327-350
Method: X-RAY DIFFRACTION / Resolution: 1.75 Å

PDB-6n85:
Resistance to inhibitors of cholinesterase 8A (Ric8A) protein in complex with MBP-tagged transducin-alpha residues 327-350
Method: X-RAY DIFFRACTION / Resolution: 2.5 Å

PDB-6n86:
Resistance to inhibitors of cholinesterase 8A (Ric8A) protein
Method: X-RAY DIFFRACTION / Resolution: 3.9 Å

Chemicals

ChemComp-SO4:
SULFATE ION

ChemComp-HOH:
WATER

Source
  • bos taurus (cattle)
  • escherichia coli o157:h7 (bacteria)
  • homo sapiens (human)
KeywordsCHAPERONE / G alpha / MBP / Ric8a / Armadillo repeat

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