High-affinity engineered chromodomains
AssigneeAxoiya Inc.
InventorsSachdev S. SIDHU, Donald Gregory MARTYN, Gianluca VEGGIANI
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Abstract
The present disclosure provides for engineered chromodomains for enhanced binding affinity to methylated proteins, their use in screening, gene transcription regulation, and in methods of their administration. The compositions and methods provided herein can be used for targeting protein methylation as a means for identifying markers of pathology for diagnostic or therapeutic applications.
Claims24
An engineered chromodomain comprising:
- a methyllysine-binding pocket, wherein the engineered chromodomain comprises at least one amino acid substitution in at least one of two regions that line the methyllysine-binding pocket as compared to a reference chromodomain selected from any one of SEQ ID NOS:
- 21-28, wherein a first region comprises amino acid positions 8-28 of the reference chromodomain, wherein a second region comprises amino acid positions 33-52 of the reference chromodomain, wherein the engineered chromodomain provides for enhanced binding to at least one methylated protein as compared to said reference chromodomain, wherein the at least one amino acid substitution comprises substitution of a neutral or positively charged amino acid residue to a negatively charged amino acid residue.
A composition comprising an engineered chromodomain comprising:
- a methyllysine-binding pocket, wherein the engineered chromodomain comprises at least one amino acid substitution in at least one of two regions that line the methyllysine-binding pocket as compared to a reference chromodomain selected from any one of SEQ ID NOS:
- 21-28, wherein a first region comprises amino acid positions 8-28 of the reference chromodomain, wherein a second region comprises amino acid positions 33-52 of the reference chromodomain, wherein the engineered chromodomain provides for enhanced binding to at least one methylated protein as compared to said reference chromodomain, wherein the at least one amino acid substitution comprises substitution of a neutral or positively charged amino acid residue to a negatively charged amino acid residue;
- and
- a buffer.
A kit comprising an engineered chromodomain comprising an amino acid sequence having at least 99% sequence identity to an amino acid sequence selected from SEQ ID NOS:
- 29-30 or SEQ ID NOS:
- 32-36, or at least 95% sequence identity to the amino acid sequence of SEQ ID NO:
- 31.
A method of detecting presence of a methylated protein in a biological sample comprising:
- (a) providing a biological sample;
- (b) contacting the biological sample with an engineered chromodomain that comprises a methyllysine-binding pocket, wherein the engineered chromodomain comprises at least one amino acid substitution in at least one of two regions that line the methyllysine-binding pocket as compared to a reference chromodomain selected from any one of SEQ ID NOS:
- 21-28, wherein a first region comprises amino acid positions 8-28 of the reference chromodomain, wherein a second region comprises amino acid positions 33-52 of the reference chromodomain, wherein the engineered chromodomain provides for enhanced binding to at least one methylated protein as compared to said reference chromodomain, wherein the at least one amino acid substitution comprises substitution of a neutral or positively charged amino acid residue to a negatively charged amino acid residue;
- and
- (c) detecting interaction of the biological sample with the engineered chromodomain, wherein the interaction indicates the presence of a methylated protein in the biological sample.
Dependencies are inferred from the claim text.