Open AccessRefolding, crystallization and preliminary X‐ray crystallographic study of the whole extracellular regions of nectins
Author(s) -
Narita Hirotaka,
Nakagawa Atsushi,
Yamamoto Yasunori,
Sakisaka Toshiaki,
Takai Yoshimi,
Suzuki Mamoru
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430911100337x
Subject(s) - nectin , adherens junction , extracellular , crystallization , cadherin , chemistry , cell adhesion molecule , adhesion , molecule , crystallography , cell adhesion , biology , biochemistry , cell , microbiology and biotechnology , organic chemistry
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 P 2 1 3 and the nectin‐2‐EC crystals belonged to space group P 6 1 22 or P 6 5 22.