SASDF65: Resistance to inhibitors of cholinesterase 8 homolog A (Ric8a) am...

Basic information

• Entry: Database: SASBDB / ID: SASDF65

Sample

Resistance to inhibitors of cholinesterase 8 homolog A (Ric8a) amino acids 1-492

• Resistance to inhibitors of cholinesterase 8 homolog A (protein), Ric8a, Bos taurus

Function / homology

Function:

G-protein alpha-subunit binding / guanyl-nucleotide exchange factor activity / G protein-coupled receptor signaling pathway / plasma membrane / cytoplasm

Domain/homology:

Synembryn / Guanine nucleotide exchange factor, Ric8 / Guanine nucleotide exchange factor synembryn / Armadillo-like helical / Armadillo-type fold

Component:

Synembryn-A

• Biological species: Bos taurus (cattle)
• Citation: Journal: Nat Commun / Year: 2019; Title: Structural underpinnings of Ric8A function as a G-protein α-subunit chaperone and guanine-nucleotide exchange factor.; Authors: Dhiraj Srivastava / Lokesh Gakhar / Nikolai O Artemyev / ; Abstract: 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.

Contact author

• Dhiraj Srivastava (University of Iowa, Iowa, USA)

Models

• Model #3128: ; Type: atomic / Comment: One state model / Chi-square value: 1.50747614912; Search similar-shape structures of this assembly by Omokage search (details)
• Model #3129: ; Type: atomic / Comment: Fractional contribution in X-ray scattering: 59% / Chi-square value: 1.19852188057; Search similar-shape structures of this assembly by Omokage search (details)
• Model #3130: ; Type: atomic / Comment: Fractional contribution in X-ray scattering: 29% / Chi-square value: 1.19852188057; Search similar-shape structures of this assembly by Omokage search (details)
• Model #3131: ; Type: atomic / Comment: Fractional contribution in X-ray scattering: 12% / Chi-square value: 1.19852188057; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Resistance to inhibitors of cholinesterase 8 homolog A (Ric8a) amino acids 1-492; Specimen concentration: 10 mg/ml
• Buffer: Name: 20 mM Tris, 150 mM KCl, 5 % glycerol, 1 mM TCEP / pH: 8

Entity #1575

Name: Ric8a / Type: protein
Description: Resistance to inhibitors of cholinesterase 8 homolog A
Formula weight: 55.643 / Num. of mol.: 1 / Source: Bos taurus / References: UniProt: Q5E9J8
Sequence:

GHMADPRAVA DALETGEEDV VMEALRAYNR ENSQSFTFDD AQQEDRKRLA KLLVSVLEQG LPPSRRVIWL QSIRILSRDR SCLDSFTSRR SLQALACYAG ISASQGSVPE PLNMDVVLES LKCLCNLVLS SPVAQALAAE AGLVVRLAER VGLCRQSSFP HDVQFFDLRL LFLLTALRTD VRQQLFQELQ GVRLLTRALE LTLGMTEGER HPELLPPQET ERAMEILKVL FNITFDSIKR EVDEEDAALY RHLGTLLRHC VMLAAAGDRT EELHGHAVNL LGNLPVKCLD VLLTLEPHEG SLEFLGVNMD VIRVLLSFME KRLHQTHRLK ESVAPVLSVL TECARMHRPA RKFLKAQVLP PLRDVRTRPE VGELLRNKLV RLMTHLDTDV KRVAAEFLFV LCSESVPRFI KYTGYGNAAG LLAARGLMAG GRPEGQYSED EDTDTDEYKE AKASINPVTG RVEEKPPNPM EGMTEEQKEH EAMKLVNMFD KLSRH

Experimental information

• Beam: Instrument name: Advanced Photon Source (APS), Argonne National Laboratory BioCAT 18ID; City: Lemont, IL / 国: USA / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.1033 Å / Dist. spec. to detc.: 3.5 mm
• Detector: Name: Pilatus3 X 1M / Pixsize x: 0.172 mm

Scan

Title: Resistance to inhibitors of cholinesterase 8 homolog A (Ric8a) amino acids 1-492
Measurement date: Oct 27, 2018 / Storage temperature: 4 °C / Cell temperature: 25 °C / Exposure time: 0.5 sec. / Unit: 1/A /

	Min	Max
Q	0.0054	0.3813

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 750 /

	Min	Max
Q	0.0126276	0.237683
P(R) point	1	750
R	0	127

Result

Type of curve: sec /

	Experimental	Porod
MW	64.6 kDa	55 kDa
Volume	-	88 nm3

	P(R)	Guinier	Guinier error
Forward scattering, I0	0.002138	0.00212099	2.0E-6
Radius of gyration, Rg	3.5 nm	3.37 nm	0.1

	Min	Max
D	-	12.7
Guinier point	25	111