Crystallization and X-ray diffraction analysis of human CLEC-2

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Aleksandra A Watson, C A O'Callaghan

Abstract

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.

References

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Citations

Apr 9, 2008·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Kristof Van HeckeLuc Van Meervelt
Nov 10, 2011·Toxins·Aleksandra A Watson, Christopher A O'Callaghan
May 18, 2011·Protein Expression and Purification·Aleksandra A WatsonChristopher A O'Callaghan
Jun 28, 2008·Protein Science : a Publication of the Protein Society·Aleksandra A WatsonChris A O'Callaghan
Dec 19, 2008·Current Opinion in Pharmacology·Chris A O'Callaghan
Apr 4, 2014·Cancer Metastasis Reviews·David G MenterKenneth V Honn
May 14, 2011·The Journal of Biological Chemistry·Aleksandra A WatsonChristopher A O'Callaghan
Jul 22, 2017·Cancer Metastasis Reviews·David G MenterKenneth V Honn
Jan 25, 2008·The Biochemical Journal·Charita M ChristouChris A O'Callaghan
Nov 30, 2006·The Journal of Biological Chemistry·Aleksandra A WatsonChristopher A O'Callaghan
Oct 15, 2009·Biochemistry·Aleksandra A WatsonChris A O'Callaghan

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