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.

• Keywords
• crystal structure, endolysin, histidine tag, lysozyme, phage T4, protein purification,
• MeSH
• Bacteriophage T4 * enzymology   genetics   MeSH
• Chromatography, Affinity methods   MeSH
• Crystallography, X-Ray methods   MeSH
• Muramidase * chemistry   genetics   isolation & purification   MeSH
• Mutation * MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Muramidase * MeSH