TY - JOUR
T1 - RGK family G-domain:GTP analog complex structures and nucleotide-binding properties
AU - Sasson, Yehezkel
AU - Navon-Perry, Leehee
AU - Huppert, Dan
AU - Hirsch, Joel A.
N1 - Funding Information:
We wish to thank Tally Shwartzman, Neta Tobis, and Orna Chomsky-Hecht for assistance with mutagenesis and subcloning and Moshe Dessau, Lior Almagor, Dana Sachyani, and the staff at ESRF for help with data collection. We are indebted to Lior Almagor and Oshik Segev for advice on fluorescence spectroscopy. Thanks are due to Yael Litvak for execution of the GTP hydrolysis assay. We thank Drs. Kathy Kelly and Geoff Pitt for critical reading of the manuscript. This study was supported in part by Israel Cancer Research Fund and Deutsche Israel Programme grants to J.A.H.
PY - 2011/10/21
Y1 - 2011/10/21
N2 - The RGK family of small G-proteins, including Rad, Gem, Rem1, and Rem2, is inducibly expressed in various mammalian tissues and interacts with voltage-dependent calcium channels and Rho kinase. Many questions remain regarding their physiological roles and molecular mechanism. Previous crystallographic studies reported RGK G-domain:guanosine di-phosphate structures. To test whether RGK proteins undergo a nucleotide-induced conformational change, we determined the crystallographic structures of Rad:GppNHp and Rem2:GppNHp to 1.7 and 1.8 Å resolutions, respectively. Also, we characterized the nucleotide-binding properties and conformations for Gem, Rad, and several structure-based mutants using fluorescence spectroscopy. The results suggest that RGK G-proteins may not behave as Ras-like canonical nucleotide-induced molecular switches. Further, the RGK proteins have differing structures and nucleotide-binding properties, which may have implications for their varied action on effectors.
AB - The RGK family of small G-proteins, including Rad, Gem, Rem1, and Rem2, is inducibly expressed in various mammalian tissues and interacts with voltage-dependent calcium channels and Rho kinase. Many questions remain regarding their physiological roles and molecular mechanism. Previous crystallographic studies reported RGK G-domain:guanosine di-phosphate structures. To test whether RGK proteins undergo a nucleotide-induced conformational change, we determined the crystallographic structures of Rad:GppNHp and Rem2:GppNHp to 1.7 and 1.8 Å resolutions, respectively. Also, we characterized the nucleotide-binding properties and conformations for Gem, Rad, and several structure-based mutants using fluorescence spectroscopy. The results suggest that RGK G-proteins may not behave as Ras-like canonical nucleotide-induced molecular switches. Further, the RGK proteins have differing structures and nucleotide-binding properties, which may have implications for their varied action on effectors.
KW - G-protein
KW - conformational switch
KW - crystallography
KW - fluorescence spectroscopy
UR - http://www.scopus.com/inward/record.url?scp=80054729017&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2011.08.017
DO - 10.1016/j.jmb.2011.08.017
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AN - SCOPUS:80054729017
SN - 0022-2836
VL - 413
SP - 372
EP - 389
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 2
ER -