Clostridial neurotoxins comprising an exogenous activation loop

Abstract

An engineered clostridial neurotoxin comprising an activation loop that comprises an amino acid sequence of the formula Cys-(Xaa)a-Ile-Asp/Glu-Gly-Arg-(Yaa)b-Cys(SEQ ID NO: 1), wherein a is an integer from 1 to 10, b is an integer from 4 to 15, each iteration of Xaa and Yaa individually represents an amino acid, and the engineered clostridial neurotoxin is not BoNT/C1, a method for producing the same, a method of treating a disease or condition comprising administering the engineered clostridial neurotoxin or the corresponding di-chain clostridial neurotoxin, a composition comprising the engineered clostridial neurotoxin or the corresponding di-chain clostridial neurotoxin, and a polynucleotide encoding the engineered clostridial neurotoxin. A method for proteolytically processing a single-chain clostridial neurotoxin into a corresponding di-chain clostridial neurotoxin, the method comprising contacting the single-chain clostridial neurotoxin with enterokinase or factor Xa and a di-chain clostridial neurotoxin produced using such a method. A method for hydrolyzing a peptide bond of a polypeptide comprising contacting the polypeptide with an enterokinase.

Dependencies are inferred from the claim text.