Method of Making Cyclic Polypeptides with Inteins
OwnerAlexander R. Horswillinventor
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Abstract
Methods for producing cyclic polypeptides comprising a lactone or thiolactone ring. In certain cases, intein fusion proteins may be used in a method for biologically producing internally cyclized polypeptides. The new methods enable the construction of cyclic polypeptide libraries that may be screened for the biological activity of cyclic polypeptides. Methods provided may be used, for instance, to produce and optimize novel cyclic peptides for use in treating Staphylococcal infections.
Claims45
A method for producing at least a first internally cyclized polypeptide comprising:
- a) expressing at least a first fusion protein in a cell, wherein the fusion protein comprises (i) a functional intein polypeptide fused to the carboxy-terminus of (ii) a polypeptide domain having at least one internal cysteine or serine residue;
- and
- b) exposing at least a first fusion protein to reducing conditions, thereby producing an internally cyclized polypeptide and a free intein polypeptide.
The method of claim 6, wherein the purification tag is a chitin binding domain (CBD).
Dependencies are inferred from the claim text.