Crystallization and X-ray diffraction analysis of human CLEC-2.
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 A using in-house radiation (lambda = 1.5418 A). These crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 35.407, b = 55.143, c = 56.078 A. The presence of one molecule per asymmetric unit is consistent with a crystal volume per unit weight (VM) of 1.82 A3 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.
Watson
Aleksandra A
AA
Henry Wellcome Building of Molecular Physiology, University of Oxford, Oxford OX3 7BN, England.
O'Callaghan
Christopher A
CA
D001792:Blood Platelets / Q000378:metabolism; D003001:Cloning, Molecular; D003460:Crystallization; D003461:Crystallography; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013927:Thrombosis / Q000378:metabolism; D014757:Viper Venoms / Q000737:chemistry; D014961:X-Ray Diffraction
C104778:CLEC2B protein, human; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D012997:Solvents; D014757:Viper Venoms; C112292:rhodocytin protein, Calloselasma rhodostoma; D011092:Polyethylene Glycols