Crystallization of rat liver macrophage migration inhibitory factor for MAD analysis.
The recombinant macrophage migration inhibitory factor (MIF) of a rat liver was crystallized using the hanging-drop vapor diffusion method. We also crystallized a selenomethionyl rat MIF under similar conditions for X-ray structure analysis using the multiwavelength anomalous diffraction method. Furthermore, two kinds of selenomethionyl rat MIF mutants, in which one or two of three methionine residues were replaced by alanine [(Met101Ala), (Met2Ala and Met101Ala)], were constructed and crystallized. These crystals belong to two different forms of the hexagonal space group P6(3). The first form has cell dimensions of a = b = 95.7 A and c = 87.9 A and contains two dimers in an asymmetric unit, while the second form, whose cell dimensions are a = b = 61.8 A and c = 53.4 A, contains only one subunit in an asymmetric unit. Preliminary X-ray examination shows that these crystals are suitable for high-resolution structure analysis.
Sugimoto
H
H
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
Suzuki
M
M
Nakagawa
A
A
Tanaka
I
I
Fujinaga
M
M
Nishihira
J
J
D000818:Animals; D001483:Base Sequence; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D008263:Macrophage Migration-Inhibitory Factors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
D008263:Macrophage Migration-Inhibitory Factors; D011993:Recombinant Fusion Proteins