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US 2010/0136535 A1Published application
Means and methods for detecting protein-peptide interactions
AssigneeATIFTUNG CAESAR CENTER OF ADVANCED EUROPEAN STUDIE AND Research
InventorsHanjo Hennemann, Claudia Kruse, Sabine Hanke, Annette Friebe
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Abstract
A novel method of detecting and characterizing protein-peptide interactions is provided, which can be used for isolating peptides and proteins, respectively, as well as in drug screening and development.
Claims20
A method of identifying a protein-peptide interaction or isolating a partner of a protein-peptide interaction, comprising:
- (a) providing with a host cell
- (i) a hybrid molecule, comprising a target protein and an effector;
- (ii) a peptide or a population of peptides, wherein the peptide is linked to a carrier protein, said carrier protein being linked to a cell compartment localisation domain, wherein said carrier protein is selected from the group consisting of thioredoxin, thioredoxin-like molecule, green fluorescent protein, Z-domain of protein A, tendamistat, Kunitz domain, fibronectin type III domain, lipocalin, bais prancreatic trypsin inhibitor and conotoxine, and
- (b) detection of a signal identifying the protein-peptide interaction of the hybrid molecule and the peptide;
- and optionally
- (c) isolating a partner of the protein-peptide interaction based on its ability to alter the signal when present.
3 more elements
A method of detecting and/or isolating an interacting peptide in a population of peptides comprising:
- (a) providing a hybrid molecule comprising a target protein and an effector within a host cell;
- (b) introducing a test peptide or a population of test peptides into the host cell, wherein the peptide is linked to a carrier protein, said carrier protein being linked to a cell compartment localization domain, wherein said carrier protein is selected from the group consisting of thioredoxin, thioredoxin-like molecule, green fluorescent protein, Z-domain of protein A, tendamistat, Kunitz domain, fibronectin type III domain, lipocalin, bais prancreatic trypsin inhibitor and conotoxine, and
- (c) detection of a signal identifying the protein-peptide interaction of the hybrid molecule and the peptide;
- and optionally
- (d) isolating a peptide, identified as a binding partner based on its ability to alter the signal when present.
2 more elements
A method of isolating a protein comprising:
- (a) providing a host cell comprising a test protein or a population of test proteins and an effector, or introducing the effector;
- (b) introducing the linkage of the effector to the test protein or population of test proteins;
- (c) providing or introducing a peptide or population of peptides within/into the host cell, wherein the peptide is linked to a carrier protein, said carrier protein being linked to a cell compartment localization domain, wherein said carrier protein is selected from the group consisting of thioredoxin, thioredoxin-like molecule, green fluorescent protein, Z-domain of protein A, tendamistat, Kunitz domain, fibronectin type III domain, lipocalin, bais prancreatic trypsin inhibitor and conotoxine, and
- (d) detection of a signal identifying the protein-peptide interaction;
- and
- (e) isolating the protein identified as a binding partner based on its ability to alter the signal when present.
3 more elements
A peptide essentially consisting essentially of SEQ ID NO:
- 6, SEQ ID NO:
- 8 or SEQ ID NO:
- 10.
Dependencies are inferred from the claim text.