Covalent tethering of functional groups to proteins and substrates therefor

摘要

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.

主权项

1. A fusion polypeptide of a protein of interest and a mutant haloalkane dehalogenase having at least 85% sequence identity to the wild type dehalogenase polypeptide of SEQ ID NO:82 and comprising one to three amino acid substitutions at positions 106, 130, and 272 relative to the wild-type dehalogenase.