Expression, purification, and preliminary X-ray crystallographic analysis of the complex of Gαi3-RGS5 from human with GDP/Mg 2+/AlF4-

Kyung Hee Rhee; Ki Hyun Nam; Won Ho Lee; Young Gyu Ko; Eunkyeong Kim Eunice; Yeon Hwang Kwang

doi:10.2174/092986606778256225

Expression, purification, and preliminary X-ray crystallographic analysis of the complex of G_αi3-RGS5 from human with GDP/Mg ²⁺/AlF₄^-

Kyung Hee Rhee, Ki Hyun Nam, Won Ho Lee, Young Gyu Ko, Eunkyeong Kim Eunice, Yeon Hwang Kwang

Research output: Contribution to journal › Article › peer-review

Abstract

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 G_i₃ from human in Escherichia coli and crystallized the complex of RGS5 and Gαi₃ proteins with GDP/Mg²⁺/AlF₄^- at 3.0 Å resolution using a synchrotron radiation source. The complex crystals belong to the tetragonal space group P4₁2₁2 or P4₃2 ₁2, with unit cell parameters a=b=95.9 Å, and c=138.8 Å. Assuming one complex protein in the crystallographic asymmetric unit, the calculated Matthews parameter (V_M) is 2.57 Å³/Da and solvent content is 52.2%.

Original language	English
Pages (from-to)	945-949
Number of pages	5
Journal	Protein and Peptide Letters
Volume	13
Issue number	9
DOIs	https://doi.org/10.2174/092986606778256225
Publication status	Published - 2006

Keywords

Crystallization
G protein signaling pathway
Gαi
Regulator of G-protein signaling 5

ASJC Scopus subject areas

Structural Biology
Biochemistry

Access to Document

10.2174/092986606778256225

Cite this

Expression, purification, and preliminary X-ray crystallographic analysis of the complex of G_αi3-RGS5 from human with GDP/Mg ²⁺/AlF₄^-. / Rhee, Kyung Hee; Nam, Ki Hyun; Lee, Won Ho; Ko, Young Gyu; Eunice, Eunkyeong Kim; Kwang, Yeon Hwang.

In: Protein and Peptide Letters, Vol. 13, No. 9, 2006, p. 945-949.

Research output: Contribution to journal › Article › peer-review

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abstract = "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 G_i3 from human in Escherichia coli and crystallized the complex of RGS5 and Gαi3 proteins with GDP/Mg2+/AlF4- at 3.0 {\AA} resolution using a synchrotron radiation source. The complex crystals belong to the tetragonal space group P41212 or P432 12, with unit cell parameters a=b=95.9 {\AA}, and c=138.8 {\AA}. Assuming one complex protein in the crystallographic asymmetric unit, the calculated Matthews parameter (VM) is 2.57 {\AA}3/Da and solvent content is 52.2%.",

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AU - Rhee, Kyung Hee

AU - Nam, Ki Hyun

AU - Lee, Won Ho

AU - Ko, Young Gyu

AU - Eunice, Eunkyeong Kim

AU - Kwang, Yeon Hwang

PY - 2006

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N2 - 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 G_i3 from human in Escherichia coli and crystallized the complex of RGS5 and Gαi3 proteins with GDP/Mg2+/AlF4- at 3.0 Å resolution using a synchrotron radiation source. The complex crystals belong to the tetragonal space group P41212 or P432 12, with unit cell parameters a=b=95.9 Å, and c=138.8 Å. Assuming one complex protein in the crystallographic asymmetric unit, the calculated Matthews parameter (VM) is 2.57 Å3/Da and solvent content is 52.2%.

AB - 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 G_i3 from human in Escherichia coli and crystallized the complex of RGS5 and Gαi3 proteins with GDP/Mg2+/AlF4- at 3.0 Å resolution using a synchrotron radiation source. The complex crystals belong to the tetragonal space group P41212 or P432 12, with unit cell parameters a=b=95.9 Å, and c=138.8 Å. Assuming one complex protein in the crystallographic asymmetric unit, the calculated Matthews parameter (VM) is 2.57 Å3/Da and solvent content is 52.2%.

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KW - Gαi

KW - Regulator of G-protein signaling 5

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