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 t...

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Bibliographic Details
Published in:Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2005-12, Vol.61 (12), p.1094-1096
Main Authors: Watson, Aleksandra A., O'Callaghan, Christopher A.
Format: Article
Language:English
Subjects:
aggretin
Blood Platelets - metabolism
C-type lectins
CLEC-2
CLEC1B
Cloning, Molecular
CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY
CRYSTALLIZATION
Crystallization Communications
Crystallography
Crystallography, X-Ray
CRYSTALS
DIFFUSION
ESCHERICHIA COLI
Escherichia coli - metabolism
Humans
Lectins, C-Type - chemistry
Membrane Glycoproteins - chemistry
MOLECULES
OPTIMIZATION
Plasmids - metabolism
platelets
Polyethylene Glycols - chemistry
Protein Folding
Protein Structure, Tertiary
RECEPTORS
Recombinant Proteins - chemistry
rhodocytin
SOLVENTS
Solvents - chemistry
SPACE GROUPS
SURFACES
thrombosis
Thrombosis - metabolism
Viper Venoms - chemistry
WEIGHT
X-RAY DIFFRACTION
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Summary: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 P212121, 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 (VM) of 1.82 Å3 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.
ISSN:1744-3091
1744-3091
DOI:10.1107/S1744309105037991