Purification, crystallization and preliminary X-ray analysis of human GIMAP2.
GTPases of immunity-associated proteins (GIMAPs) are important regulators of T-cell death and survival. Here, the crystallization and data collection of three GIMAP2 constructs in various nucleotide-loaded states is described. Selenomethionine-substituted carboxy-terminally truncated GIMAP2 (amino-acid residues 1-260; GIMAP2(1-260)) in the nucleotide-free form crystallized in space group P2(1)2(1)2(1) and the crystals diffracted X-rays to 1.5 A resolution. The phase problem was solved using the single anomalous dispersion (SAD) protocol. GDP-bound GIMAP2(21-260) and GDP-bound GIMAP2(1-234) crystallized in space group P2(1)2(1)2(1) and the crystals diffracted X-rays to 2.9 and 1.7 A resolution, respectively. GTP-bound GIMAP2(1-234) crystallized in space group C222(1) and the crystals diffracted to 1.9 A resolution. These results will allow a detailed structural analysis of GIMAP2, which will provide insight into the architecture and function of the GIMAP family.
Schwefel
David
D
Max-Delbrück-Centrum für Molekulare Medizin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany. david.schwefel@mdc-berlin.de
Fröhlich
Chris
C
Daumke
Oliver
O
D003460:Crystallization; D018360:Crystallography, X-Ray; D020558:GTP Phosphohydrolases / Q000737:chemistry*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*
D008565:Membrane Proteins; C553225:GIMAP2 protein, human; D020558:GTP Phosphohydrolases