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TitleStructure, Function, and Dynamics of the Gα Binding Domain of Ric-8A.
Journal, issue, pagesStructure, Vol. 27, Issue 7, Page 1137-11147.e5, Year 2019
Publish dateJul 2, 2019
AuthorsBaisen Zeng / Tung-Chung Mou / Tzanko I Doukov / Andrea Steiner / Wenxi Yu / Makaia Papasergi-Scott / Gregory G Tall / Franz Hagn / Stephen R Sprang /
PubMed AbstractRic-8A is a 530-amino acid cytoplasmic molecular chaperone and guanine nucleotide exchange factor (GEF) for i, q, and 12/13 classes of heterortrimeric G protein alpha subunits (Gα). We report the 2. ...Ric-8A is a 530-amino acid cytoplasmic molecular chaperone and guanine nucleotide exchange factor (GEF) for i, q, and 12/13 classes of heterortrimeric G protein alpha subunits (Gα). We report the 2.2-Å crystal structure of the Ric-8A Gα-binding domain with GEF activity, residues 1-452, and is phosphorylated at Ser435 and Thr440. Residues 1-429 adopt a superhelical fold comprised of Armadillo (ARM) and HEAT repeats, and the C terminus is disordered. One of the phosphorylated residues potentially binds to a basic cluster in an ARM motif. Amino acid sequence conservation and published hydrogen-deuterium exchange data indicate repeats 3 through 6 to be a putative Gα-binding surface. Normal mode modeling of small-angle X-ray scattering data indicates that phosphorylation induces relative rotation between repeats 1-4, 5-6, and 7-9. 2D H-N-TROSY spectra of [H,N]-labeled Gαi1 in the presence of R452 reveals chemical shift perturbations of the C terminus and Gαi1 residues involved in nucleotide binding.
External linksStructure / PubMed:31155309 / PubMed Central
MethodsSAS (X-ray synchrotron) / X-ray diffraction
Resolution2.2 - 2.3 Å
Structure data

SASDFA5: Unposphorylated Resistance to inhibitors of cholinesterase 8 homolog A, Ric-8A, amino acids 1-452 (Rattus norvegicus)
Method: SAXS/SANS

SASDFB5: Phosphorylated Resistance to inhibitors of cholinesterase 8 homolog A, Ric-8A, amino acids 1-452 (Rattus norvegicus)
Method: SAXS/SANS

PDB-6nmg:
Crystal Structure of Rat Ric-8A G alpha binding domain
Method: X-RAY DIFFRACTION / Resolution: 2.2 Å

PDB-6nmj:
Crystal Structure of Rat Ric-8A G alpha binding domain, "Paratone-N Immersed"
Method: X-RAY DIFFRACTION / Resolution: 2.3 Å

Chemicals

ChemComp-SO4:
SULFATE ION

ChemComp-HOH:
WATER

Source
  • rattus norvegicus (Norway rat)
KeywordsSIGNALING PROTEIN / guanine nucleotide exchange factor heterotrimeric G protein alpha subunit G alpha i(1)

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