1f3b4efedd The complex formation efficiency (%) at 4 C and complex stability (%) at 36.3 C were defined and determined as described in methods section. Maeda, S. The formed Rho*–Gi complex was visualized by native gel electrophoresis and the gel bands of complex were quantified by ImageJ software. Full size image Enable zoom Nature Structural & Molecular Biology ISSN: 1545-9993 EISSN: 1545-9985 About NPG Contact NPG Accessibility statement Help Privacy policy Use of cookies Legal notice Terms Nature jobs Nature Asia Nature Education RSS web feeds Search:Go 2015 Macmillan Publishers Limited, part of Springer Nature. The complex formation efficiency (%) at 4 C and complex stability (%) at 36.3 C were defined and determined as described in methods section. Full size image Enable zoom Supplementary Figure 7: Structural overlay of Gαi1–GDP with G protein α subunit from the plant Arabidopsis thaliana (AtGPA1). From Probing Gαi1 protein activation at single–amino acid resolution Dawei Sun1, 2, Tilman Flock3, n2 Xavier Deupi1, 4, n2 Shoji Maeda1, n1 Milos Matkovic1, 2, Sandro Mendieta1, Daniel Mayer1, 2, Roger J P Dawson5, Gebhard F X Schertler1, 2, M Madan Babu3, Dmitry B Veprintsev1, 2, Journal name: Nature Structural & Molecular Biology Volume: 22, Pages: 686–694 Year published: (2015) DOI: doi:10.1038/nsmb.3070 Received 20 April 2015 Accepted 17 July 2015 Published online 10 August 2015 a, Visualization of Rho*–Gi complex by the native gel electrophoresis. The thermal stability measurement of all Gαi1 alanine mutants in the nucleotide-bound state was performed upon addition of 1mM GDP or 100 M GTPγS. c, Tm of Gαi1 alanine mutants which stabilize the receptor-bound state. contributed to the discussion and to writing the manuscript.
and D.B.V. Previous Figures index Next Additional data Author footnotes Present address: Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA.Shoji MaedaThese authors contributed equally to this work.Tilman Flock & Xavier Deupi Affiliations Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland. Full size image Enable zoom Supplementary Figure 8: Mutations that affect the GTP-bound state cluster around the γ-phosphate of the GTPγS. The determined Td50 value of Rho*–Gi(WT) is 36.0 0.1 C. b, Gαi1 alanine mutants stabilize the Rho*–Gi complex. The Rho*–Gi complexes were formed with WT, Y320AG.S6.2, L348AG.H5.20, G352AG.H5.24 and L353AG.H5.25 of Gαi1 at 4 C and heated at 36.3 C, followed by the native gel electrophoresis. D.S., X.D. e, Thermal profile of F336AG.H5.8 in the nucleotide-bond state. The structure of AtGPA1-GTPγS (PDB 2XTZ32) is shown in light pink. The results clearly show that these four Gαi1 alanine mutants are severely impaired in their ability to form the Rho*–Gi complex.
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