Expression, crystallization and preliminary X-ray diffraction analysis of human paired Ig-like type 2 receptor alpha (PILRalpha).
Human paired immunoglobulin-like (Ig-like) type 2 receptor alpha (PILRalpha) is a type I membrane protein that is mainly expressed in immune-related cells such as monocytes, granulocytes and dendritic cells. PILRalpha can suppress the functions of such immune cells because it has the immunoreceptor tyrosine-based inhibitory motif (ITIM) in the intracellular region, which recruits the phosphatase Src homology-2 (SH2) domain-containing protein tyrosine phosphatase 2 (SHP-2) to inhibit phosphorylations induced by activation signals. The extracellular region of human PILRalpha comprises one immunoglobulin superfamily V-set domain and a stalk region. The V-set domain (residues 13-131) of human PILRalpha was overexpressed in Escherichia coli as inclusion bodies, refolded by rapid dilution and purified. The PILRalpha protein was successfully crystallized at 293 K using the sitting-drop vapour-diffusion method. The crystals diffracted to 1.3 A resolution at SPring-8 BL41XU; they belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 40.4, b = 45.0, c = 56.9 A, and contain one molecule per asymmetric unit.
Tabata
Shigekazu
S
Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Kuroki
Kimiko
K
Maita
Nobuo
N
Wang
Jing
J
Shiratori
Ikuo
I
Arase
Hisashi
H
Kohda
Daisuke
D
Maenaka
Katsumi
K
D002850:Chromatography, Gel; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D017931:DNA Primers; D003713:Dendritic Cells / Q000276:immunology; D006098:Granulocytes / Q000276:immunology; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D009000:Monocytes / Q000276:immunology; D011971:Receptors, Immunologic / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
D017931:DNA Primers; D008562:Membrane Glycoproteins; C412263:PILRA protein, human; D011971:Receptors, Immunologic; D011994:Recombinant Proteins