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Yorodumi- SASDGB6: Resistance to inhibitors of cholinesterase 8 homolog A (Ric8A) mi... -
+Open data
-Basic information
Entry | Database: SASBDB / ID: SASDGB6 |
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Sample | Resistance to inhibitors of cholinesterase 8 homolog A (Ric8A) miniGi complex
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Function / homology | Function and homology information G-protein alpha-subunit binding / guanyl-nucleotide exchange factor activity / G protein-coupled receptor signaling pathway / plasma membrane / cytoplasm Similarity search - Function |
Biological species | Bos taurus (cattle) synthetic construct (others) |
Citation | Journal: J Biol Chem / Year: 2019 Title: Large-scale conformational rearrangement of the α5-helix of Gα subunits in complex with the guanine nucleotide exchange factor Ric8A. Authors: Dhiraj Srivastava / Nikolai O Artemyev / Abstract: Resistance to inhibitors of cholinesterase 8A (Ric8A) protein is an important G protein-coupled receptor (GPCR)-independent regulator of G protein α-subunits (Gα), acting as a guanine nucleotide ...Resistance to inhibitors of cholinesterase 8A (Ric8A) protein is an important G protein-coupled receptor (GPCR)-independent regulator of G protein α-subunits (Gα), acting as a guanine nucleotide exchange factor (GEF) and a chaperone. Insights into the complex between Ric8A and Gα hold the key to understanding the mechanisms underlying noncanonical activation of G-protein signaling as well as the folding of nascent Gα proteins. Here, we examined the structure of the complex of Ric8A with minimized Gα (miniGα) in solution by small-angle X-ray scattering (SAXS) and exploited the scattering profile in modeling of the Ric8A/miniGα complex by steered molecular dynamics (SMD) simulations. A small set of models of the complex featured minimal clash scores, excellent agreement with the experimental SAXS data, and a large-scale rearrangement of the signal-transducing α5-helix of Gα away from its β-sheet core. The resulting interface involved the Gα α5-helix bound to the concave surface of Ric8A and the Gα β-sheet that wraps around the C-terminal part of the Ric8A armadillo domain, leading to a severe disruption of the GDP-binding site. Further modeling of the flexible C-terminal tail of Ric8A indicated that it interacts with the effector surface of Gα. This smaller interface may enable the Ric8A-bound Gα to interact with GTP. The two-interface interaction with Gα described here distinguishes Ric8A from GPCRs and non-GPCR regulators of G-protein signaling. |
Contact author |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Models
Model #3814 | Type: atomic / Chi-square value: 1.637 Search similar-shape structures of this assembly by Omokage search (details) |
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Model #3813 | Type: atomic / Software: (Rosetta) / Chi-square value: 1.882 Search similar-shape structures of this assembly by Omokage search (details) |
-Sample
Sample | Name: Resistance to inhibitors of cholinesterase 8 homolog A (Ric8A) miniGi complex Specimen concentration: 10 mg/ml / Entity id: 1575 / 1912 |
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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 ...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 |
Entity #1912 | Type: protein / Description: miniGi / Formula weight: 24.54 / Num. of mol.: 1 / Source: synthetic construct Sequence: AMEKAAREVK LLLLGADNSG KSTIVKQMKI IHEAGEYMPM ERVKTTGIVE THFTFKDLHF KMFDVGGQRS ERKKWIHCFE DVAAIIFCVD LSDYEEMNRM HESMKLFDSI CNNKWFTDTS IILFLNKKDL FEEKIKKSPL TICYQEYAGS NTYEEAAAYI QCQFEDLNKR ...Sequence: AMEKAAREVK LLLLGADNSG KSTIVKQMKI IHEAGEYMPM ERVKTTGIVE THFTFKDLHF KMFDVGGQRS ERKKWIHCFE DVAAIIFCVD LSDYEEMNRM HESMKLFDSI CNNKWFTDTS IILFLNKKDL FEEKIKKSPL TICYQEYAGS NTYEEAAAYI QCQFEDLNKR KDTKEIYTHF TCATDTKNVQ FVFDAVTDVI IKNNLKDCGL F |
-Experimental information
Beam | Instrument name: Advanced Photon Source (APS), Argonne National Laboratory BioCAT 18ID City: Lemont, IL / 国: USA / Type of source: X-ray synchrotron | ||||||||||||||||||||||||
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Detector | Name: Pilatus3 X 1M / Pixsize x: 0.172 mm | ||||||||||||||||||||||||
Scan | Title: Resistance to inhibitors of cholinesterase 8 homolog A (Ric8A) miniGi complex Measurement date: Oct 27, 2018 / Storage temperature: 4 °C / Exposure time: 0.5 sec. / Unit: 1/A /
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Distance distribution function P(R) | Sofotware P(R): GNOM 4.6 / Number of points: 420 /
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Result | Experimental MW: 83.6 kDa / Type of curve: sec Comments: Purified Ric8a1-492 was mixed with excess of purified miniGi and the complex was purified by size-exclusion chromatography (SEC). The complex fractions were pooled together, concentrated ...Comments: Purified Ric8a1-492 was mixed with excess of purified miniGi and the complex was purified by size-exclusion chromatography (SEC). The complex fractions were pooled together, concentrated and additional SEC-SAXS data was collected as described above. X-ray wavelength = UNKNOWN.
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