Methods of identifying SENP1 inhibitors
Provided herein are methods of detecting binding of an SENP1 polypeptide to a compound and methods for screening for inhibitors of SENP1. Further provided are aqueous compositions comprising SENP1 polypeptides and NMR apparatuses comprising the compositions for NMR analysis.
1. A method of detecting binding of an SENP1 polypeptide to a compound, the method comprising:
(i) contacting an SENP1-SUMO complex with a compound, wherein the SENP1-SUMO complex comprises an SENP1 polypeptide and a SUMO protein;
(ii) allowing the compound to bind to the SENP1 polypeptide of the SENP1-SUMO complex, thereby forming a SENP1-SUMO-compound complex;
(iii) detecting the SENP1-SUMO-compound complex using nuclear magnetic resonance, thereby detecting binding of the SENP1 polypeptide to the compound.
2. The method of claim 1 , wherein the detecting comprises
determining a chemical shift for an amino acid in an active site of the SENP1 polypeptide.
3. The method of claim 2 , wherein the chemical shift in the presence of the compound is changed relative to the corresponding chemical shift in the absence of the compound.
4. The method of claim 3 , wherein the amino acid is selected from the group consisting of an amino acid residue corresponding to D550, H533, C603, W465, W534, L466, G531, C535, M552, G554, E469 and Q596 of SEQ ID NO: 1.
5. The method of claim 1 , wherein the SENP1 polypeptide comprises SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7.
6. The method of claim 1 , wherein the SENP1 polypeptide comprises a mutation at amino acid residue corresponding to amino acid residue 603 of SEQ ID NO: 1.
7. The method of claim 1 , wherein the detecting comprises producing an NMR spectra of the SENP1-SUMO-compound complex and identifying a change in the NMR spectra relative to the absence of the compound.
8. The method of claim 7 , wherein the change is a change in the chemical shift of an amino acid of SEQ ID NOs: 3, 4, 5, 6 or 7.
9. The method of claim 7 , wherein the change is a change in the chemical shift of the amino acid 5603.
10. The method of claim 7 , wherein the change is a change in the chemical shift of an amino acid residue 440-455, 463-473, 493-515, 529-535, 550-554, or 596-603 of SEQ ID NO: 1.