Open AccessCrystallization and X‐ray diffraction analysis of human CLEC‐2
Author(s) -
Watson Aleksandra A.,
O'Callaghan Christopher A.
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105037991
Subject(s) - orthorhombic crystal system , crystallization , polyethylene glycol , crystallography , peg ratio , chemistry , recombinant dna , solvent , c type lectin , molecule , materials science , crystal structure , lectin , biochemistry , organic chemistry , finance , economics , gene
The human C‐type lectin‐like protein CLEC‐2 has recently been shown to be expressed on the surface of platelets and to function as a receptor for the snake‐venom protein rhodocytin. The C‐type lectin‐like domain (CTLD) of CLEC‐2 was expressed in Escherichia coli , refolded and purified. Crystals of this recombinant CLEC‐2 were grown by sitting‐drop vapour diffusion using polyethylene glycol (PEG) 6000 as a precipitant. After optimization, crystals were grown which diffracted to 2.0 Å using in‐house radiation (λ = 1.5418 Å). These crystals belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 35.407, b = 55.143, c = 56.078 Å. The presence of one molecule per asymmetric unit is consistent with a crystal volume per unit weight ( V M ) of 1.82 Å 3 Da −1 and a solvent content of 32.6%. These results suggest that crystals producing diffraction of this quality will be suitable for the structural determination of human CLEC‐2.