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Metal ions‐binding T4 lysozyme as an intramolecular protein purification tag compatible with X‐ray crystallography
Author(s) -
Boura Evzen,
Baumlova Adriana,
Chalupska Dominika,
Dubankova Anna,
Klima Martin
Publication year - 2017
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.3162
Subject(s) - lysozyme , protein tag , chemistry , protein purification , protein crystallization , fusion protein , recombinant dna , maltose binding protein , metal ions in aqueous solution , crystallization , biochemistry , myc tag , affinity chromatography , crystallography , metal , enzyme , organic chemistry , gene
Phage T4 lysozyme is a well folded and highly soluble protein that is widely used as an insertion tag to improve solubility and crystallization properties of poorly behaved recombinant proteins. It has been used in the fusion protein strategy to facilitate crystallization of various proteins including multiple G protein‐coupled receptors, lipid kinases, or sterol binding proteins. Here, we present a structural and biochemical characterization of its novel, metal ions‐binding mutant (mbT4L). We demonstrate that mbT4L can be used as a purification tag in the immobilized‐metal affinity chromatography and that, in many respects, it is superior to the conventional hexahistidine tag. In addition, structural characterization of mbT4L suggests that mbT4L can be used as a purification tag compatible with X ‐ray crystallography.