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

Abstract

Recombinant human CLEC-2 was crystallized in the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1} and X-ray diffraction data were collected to 2.0 Å. 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 P2{sub 1}2{sub 1}2{sub 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{sub M}) of 1.82 Å{sup 3} Da{sup −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.

Authors:
;  [1]
  1. Henry Wellcome Building of Molecular Physiology, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)
Publication Date:
OSTI Identifier:
22356196
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 61; Journal Issue: Pt 12; Other Information: PMCID: PMC1978148; PMID: 16511244; PUBLISHER-ID: ll5044; OAI: oai:pubmedcentral.nih.gov:1978148; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; DIFFUSION; ESCHERICHIA COLI; MOLECULES; OPTIMIZATION; RECEPTORS; SOLVENTS; SPACE GROUPS; SURFACES; WEIGHT; X-RAY DIFFRACTION

Citation Formats

Watson, Aleksandra A., and O’Callaghan, Christopher A., E-mail: chrisoc@ccmp.ox.ac.uk. Crystallization and X-ray diffraction analysis of human CLEC-2. United Kingdom: N. p., 2005. Web. doi:10.1107/S1744309105037991.
Watson, Aleksandra A., & O’Callaghan, Christopher A., E-mail: chrisoc@ccmp.ox.ac.uk. Crystallization and X-ray diffraction analysis of human CLEC-2. United Kingdom. doi:10.1107/S1744309105037991.
Watson, Aleksandra A., and O’Callaghan, Christopher A., E-mail: chrisoc@ccmp.ox.ac.uk. Thu . "Crystallization and X-ray diffraction analysis of human CLEC-2". United Kingdom. doi:10.1107/S1744309105037991.
@article{osti_22356196,
title = {Crystallization and X-ray diffraction analysis of human CLEC-2},
author = {Watson, Aleksandra A. and O’Callaghan, Christopher A., E-mail: chrisoc@ccmp.ox.ac.uk},
abstractNote = {Recombinant human CLEC-2 was crystallized in the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1} and X-ray diffraction data were collected to 2.0 Å. 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 P2{sub 1}2{sub 1}2{sub 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{sub M}) of 1.82 Å{sup 3} Da{sup −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.},
doi = {10.1107/S1744309105037991},
journal = {Acta Crystallographica. Section F},
number = Pt 12,
volume = 61,
place = {United Kingdom},
year = {Thu Dec 01 00:00:00 EST 2005},
month = {Thu Dec 01 00:00:00 EST 2005}
}
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