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NECTIN蛋白胞外区域全长的重折叠、结晶及初步X射线晶体学研究

Refolding, crystallization and preliminary X-ray crystallographic study of the whole extracellular regions of nectins.

作者信息

Narita Hirotaka, Nakagawa Atsushi, Yamamoto Yasunori, Sakisaka Toshiaki, Takai Yoshimi, Suzuki Mamoru

机构信息

Laboratory of Supramolecular Crystallography, Research Center for Structural and Functional Proteomics, Institute for Protein Research, Osaka University, Suita 565-0871, Japan.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Mar 1;67(Pt 3):344-8. doi: 10.1107/S174430911100337X. Epub 2011 Feb 23.

DOI:10.1107/S174430911100337X
PMID:21393840
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3053160/
Abstract

The nectin family of Ca2+-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.

摘要

Ca2+非依赖性免疫球蛋白样细胞间粘附分子的NECTIN家族包含四个成员。NECTIN在其细胞外区域具有三个Ig样结构域，与钙粘蛋白协同形成细胞间粘附连接。NECTIN-1（NECTIN-1-EC）和NECTIN-2（NECTIN-2-EC）的整个细胞外区域在大肠杆菌中作为包涵体表达，在8M尿素中溶解，然后通过快速稀释重折叠到重折叠溶液中。随后通过三个色谱步骤纯化重折叠的蛋白质，并使用悬滴气相扩散法结晶。NECTIN-1-EC晶体属于空间群P2(1)3，NECTIN-2-EC晶体属于空间群P6(1)22或P6(5)22。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/e0ba0f190952/f-67-00344-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/423217a200d7/f-67-00344-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/697c33cdf282/f-67-00344-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/e0ba0f190952/f-67-00344-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/423217a200d7/f-67-00344-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/697c33cdf282/f-67-00344-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f53/3053160/e0ba0f190952/f-67-00344-fig3.jpg

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