Regulation of protein function by insertion of interaction peptides
AssigneeKemijski Institut National Institute of Chemistry
InventorsRoman JERALA, Tjaša PLAPER, Estera MERLJAK, Tina FINK
Life of this patent
Estimating the term…
Abstract
The invention refers to the regulation of function of proteins through insertion of a peptide into the selected protein and its interaction with a regulatory peptide that interacts with the inserted peptide. The invention can be used to activate or inactivate the function of different selected proteins and therefore to regulate their properties and processes, useful for pharmacological, therapeutic, diagnostic, sensing, biotechnological and other industrial applications.
Claims13
A method of regulating a function of a target protein comprising:
- a. genetically inserting an insertion peptide with a length of 20 to 50 amino acid residues into the selected target protein at such a position of said target protein, preferably in a solvent exposed loop, that the function of the target protein is maintained;
- b. adding a regulatory polypeptide, which specifically interacts with said inserted peptide and forms a structured dimer, in particular a heterodimeric coiled-coil with a defined structure, wherein this formation of a dimer, preferentially a coiled-coil peptide dimer, increases the distance between the amino and carboxy termini of the inserted peptide and inhibits the function of the target protein.
A method of regulating a function of a target protein, comprising:
- a. genetically inserting an insertion peptide that is by itself not structured into the selected target protein at such a position of said target protein, preferably into a solvent-exposed loop, that the function of the target protein is maintained;
- b. fusing one or two inhibitory peptide(s) that can interact with the inserted peptide to the target protein via a non-structured hydrophilic flexible linker peptide, preferentially comprising 3 to 30 amino acid residues, wherein the inhibitory peptide can bind to the inserted peptide within the same protein molecule, preferentially forming a coiled-coil dimer, thereby inhibiting the function of the target protein in a constitutively expressed state;
- wherein the function of the target protein can be regained by adding a regulatory peptide which binds tightly to the inhibitory peptide or
- by cleaving the linker between the inhibitory peptide and the target protein by a protease that has a recognition sequence included within the linker peptide.
Dependencies are inferred from the claim text.