METHOD OF IMMOBILIZING A PROTEIN OR MOLECULE VIA A MUTANT DEHALOGENASE THAT IS BOUND TO AN IMMOBILIZED DEHALOGENASE SUBSTRATE AND LINKED DIRECTLY OR INDIRECTLY TO THE PROTEIN OR MOLECULE
申请人:Promega Corporation
公开号:US20150140587A1
公开(公告)日:2015-05-21
A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
提供了一种可选地与感兴趣的蛋白质融合的突变水解酶。该突变水解酶能够与相应的非突变(野生型)水解酶的底物形成键合,该键合比野生型水解酶和底物之间形成的键合更稳定,并且相对于野生型水解酶至少有两个氨基酸替换。还提供了包含一个或多个功能基团的水解酶底物,以及使用该突变水解酶和本发明的底物的方法。还提供了一种能够与底物形成稳定键合的融合蛋白质和表达该融合蛋白质的细胞。