Expression, purification, and preliminary X-ray crystallographic analysis of the complex of G(alphai3)-RGS5 from human with GDP/Mg2+)/AlF4-.
Regulator of G-protein signaling 5 (RGS5), an inhibitor of Gq and Gi activation, is a member of the small RGS protein subfamily. However, despite significant process in the investigation of RGS5, no structure is yet available. In order to elucidate the mechanism of the RGS5 in G protein signaling pathway, we have overexpressed the RGS5 and Galphai(3) from human in Escherichia coli and crystallized the complex of RGS5 and Galphai(3) proteins with GDP/Mg(2+)/AlF(4)(-) at 3.0 A resolution using a synchrotron radiation source. The complex crystals belong to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2, with unit cell parameters a=b=95.9 A, and c=138.8 A. Assuming one complex protein in the crystallographic asymmetric unit, the calculated Matthews parameter (V(M)) is 2.57 A(3)/Da and solvent content is 52.2 %.
Rhee
Kyung-Hee
KH
Life Sciences Division, Biomedical Research Center, Korea Institute of Science and Technology, Division of Biotechnology, Korea University, Seoul 136-701, South Korea. chahong@korea.ac.kr.
Nam
Ki-Hyun
KH
Lee
Won-Ho
WH
Ko
Young-Gyu
YG
Kim
Eunice Eunkyeong
EE
Hwang
Kwang Yeon
KY
D017607:Aluminum Compounds / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D005459:Fluorides / Q000737:chemistry* / Q000378:metabolism; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008274:Magnesium / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes; D020710:RGS Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015290:Second Messenger Systems / Q000502:physiology
D017607:Aluminum Compounds; D046912:Multiprotein Complexes; D020710:RGS Proteins; C491349:RGS5 protein, human; D006153:Guanosine Diphosphate; C050992:tetrafluoroaluminate; D019204:GTP-Binding Proteins; C498819:GNAI3 protein, human; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D008274:Magnesium; D005459:Fluorides