Split modified dehalogenase variants
OwnerMichael P. Killoran, Evan Glasgow +3inventors
InventorsMichael P. Killoran, Evan Glasgow, Lance P. Encell, Thomas Kirkland, Rahele Esmatpour
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Abstract
Provided herein are peptide and polypeptide sequences that structurally assemble to form active, modified dehalogenase structures capable of binding to a haloalkyl ligand. In particular, provided herein are split dehalogenase variants that assemble through structural complementation into active dehalogenase complexes, and systems and methods of use thereof.
Claims24
1-2. (canceled)
A composition comprising a split variant of a polypeptide comprising (i) a first fragment of the polypeptide comprising at least 70% sequence identity with a first portion of SEQ ID NO:
- 1, and (ii) a second fragment of the polypeptide comprising at least 70% sequence identity with a second portion of SEQ ID NO:
- 1.
4-8. (canceled)
10-11. (canceled)
24-31. (canceled)
A polynucleotide or polynucleotides encoding the composition of claim 1.
35-43. (canceled)
A method comprising to detect a protein-protein interaction in a sample comprising contacting:
- (a) a first fusion comprising:
- (i) a first complementary fragment of a split variant of a polypeptide comprising at least 70% sequence similarity with a first portion of SEQ ID NO:
- 1;
- and
- (ii) a first protein of interest;
- (b) a second fusion comprising:
- (i) a second complementary fragment of a split variant of a polypeptide comprising at least 70% sequence similarity with a second portion of SEQ ID NO:
- 1;
- and
- (ii) a second protein of interest;
- and
- (c) a substrate comprising R-linker-A-X, wherein R is a functional group or solid support, X is a halogen, and A-X is a substrate for a dehalogenase enzyme;
- wherein binding of the first protein of interest to the second protein of interest results in formation of a complex between the first complementary fragment and the secondary complementary fragment that is capable for forming a covalent bond with the substrate.
45-46. (canceled)
Dependencies are inferred from the claim text.