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2015 ; 80
(ä): 9.10.1-9.10.37
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Protein purification using PDZ affinity chromatography
#MMPMID25829303
Walkup WG 4th
; Kennedy MB
Curr Protoc Protein Sci
2015[Apr]; 80
(ä): 9.10.1-9.10.37
PMID25829303
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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.