Protein purification using PDZ affinity chromatography.
Protein purification using PDZ affinity chromatography.
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DOI：
10.1002/0471140864.ps0910s80
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发表时间：
2015-04-01
影响因子：
--
通讯作者：
Kennedy, Mary B
中科院分区：
文献类型：
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作者：
Walkup, Ward G 4th;Kennedy, Mary B
PDZ结构域在自然界中作为支架和膜相关蛋白中的蛋白结合域发挥作用。它们由大约90个残基组成，与互补的C端肽序列、其他PDZ结构域和/或磷脂发生特异的、高亲和力的相互作用。我们之前已经证明，PDZ结构域与其配体之间的特异性强相互作用使其非常适合用于亲和层析。本单位为PDZ亲和层析程序提供了适用于纯化含有PDZ结构域或PDZ结构域结合配体的蛋白质的方案，无论是天然的还是由基因工程引入的。我们详细介绍了由PDZ结构域或PDZ结构域肽配体偶联到固体载体上的亲和树脂的制备。这些树脂可以用来纯化含有内源性或基因引入的PDZ结构域或配体的蛋白质，用游离的PDZ结构域肽配体洗脱蛋白质。©2015，John Wiley&Sons，Inc.
PDZ domains function in nature as protein‐binding domains within scaffold and membrane‐associated proteins. They comprise approximately 90 residues and undergo specific, high‐affinity interactions with complementary C‐terminal peptide sequences, other PDZ domains, and/or phospholipids. We have previously shown that the specific, strong interactions of PDZ domains with their ligands make them well suited for use in affinity chromatography. This unit provides protocols for the PDZ affinity chromatography procedure that are applicable for the purification of proteins that contain PDZ domains or PDZ domain‐binding ligands, either naturally or introduced by genetic engineering. We detail the preparation of affinity resins composed of PDZ domains or PDZ domain peptide ligands coupled to solid supports. These resins can be used to purify proteins containing endogenous or genetically introduced PDZ domains or ligands, eluting the proteins with free PDZ domain peptide ligands. © 2015 by John Wiley & Sons, Inc.