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Refolding, Crystallization and Preliminary X-ray Crystallographic Study of the Whole Extracellular Regions of Nectins

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Abstract

The nectin family of Ca(2+)-independent immunoglobulin-like cell-cell adhesion molecules contains four members. Nectins, which have three Ig-like domains in their extracellular region, form cell-cell adherens junctions cooperatively with cadherins. The whole extracellular regions of nectin-1 (nectin-1-EC) and nectin-2 (nectin-2-EC) were expressed in Escherichia coli as inclusion bodies, solubilized in 8 M urea and then refolded by rapid dilution into refolding solution. The refolded proteins were subsequently purified by three chromatographic steps and crystallized using the hanging-drop vapour-diffusion method. The nectin-1-EC crystals belonged to space group P2(1)3 and the nectin-2-EC crystals belonged to space group P6(1)22 or P6(5)22.

This article is published online in Crystallization Communications and is free to access.

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