Method for identifying inhibitors of
View Patent ↗Provided are methods for identifying and/or designing inhibitors of Staphylococcus aureus biological activity. Also provided is a computer readable medium encoded with instructions executable by a CPU for performing the functions of identifying or designing inhibitors of Staphylococcus aureus biological activity, and a system comprising the computer readable medium and a CPU. Also provided are compounds identified and/or designed by the methods, methods of treatment including the compounds, methods of inhibiting S. aureus using the compounds, medicaments including the compounds for use in the treatment of S. aureus infection, or conditions related to S. aureus infection, and pharmaceutical compositions including the compounds. Methods for identifying a bacterial cell that is resistant to methicillin are also provided.
1. A method for identifying a candidate inhibitor that preferentially binds to Staphylococcus aureus N-Succinyl-L,L-DAP Desuccinylase (DapE) or a homologue or active fragment thereof in the open conformation, comprising:
obtaining the structure coordinates of amino acids of DapE or homologue or active fragment thereof wherein the structure coordinates include the binding site of DapE or homologue or active fragment thereof comprises amino acid residues Glu-150, Ser-151, His-180, Gly-181, Glu-182, Lys-183, Arg-214, Asn-216, His-269, Arg-350, His-384, Tyr-385, Val-386, Asp-387, Lys-388, Thr-412, Gly-416, and His-440 of SEQ ID NO:2;
generating a three-dimensional model of DapE or homologue or active fragment thereof using the structure coordinates of the amino acids from DapE in closed and open conformations, wherein the model comprises a root mean square deviation from backbone atoms of said amino acids of not more than ±1.0 Å;
determining the binding site of DapE or homologue thereof in the closed confirmation from the three-dimensional model, and separately determining the binding site of DapE or homologue thereof in the open confirmation; and
performing a computer fitting analysis to identify the candidate inhibitor that interacts with the binding site of DapE in the open confirmation but does not interact with the binding site of DapE in the closed configuration, and prevents interaction of the DapE lid domain with the DapE catalytic domain, wherein the interface of the catalytic domain and the lid domain comprises amino acid residues Glu-150, Ser-151, His-180, Gly-181, Glu-182, Lys-183, Arg-214, Asn-216, His-269, Arg-350, His-384, Tyr-385, Val-386, Asp-387, Lys-388, Thr-412, Gly-416, and His-440 of SEQ ID NO:2.
2. The method of claim 1 , wherein the S. aureus DapE or active fragment thereof is from a methicillin resistant S. aureus (MRSA) strain.
3. The method for identifying a candidate inhibitor of claim 1 , further comprising contacting the identified candidate inhibitor with DapE or homologue or active fragment thereof under conditions that allow for determination the effect of the inhibitor on S. aureus biological activity.
4. The method of claim 1 , wherein the fitting analysis of the interaction between the candidate inhibitor and the binding site predicts that the inhibitor will bind at the interface between the catalytic domain and the lid domain.
5. The method of claim 1 , wherein said structure coordinates of amino acids of DapE are as found in Table S1 and Table S2.
6. A method for identifying a candidate inhibitor that preferentially binds to Staphylococcus aureus N-Succinyl-L,L-DAP Desuccinylase (DapE) or a homologue or active fragment thereof in the open conformation, comprising:
obtaining the structure coordinates of amino acids of DapE or homologue or active fragment thereof under closed (reducing) and open (oxidizing) conditions,
generating a three-dimensional model of DapE or homologue or active fragment thereof at the root mean square deviation from backbone atoms of said amino acids of not more than ±1.0 Å, using the structure coordinates of the amino acids from DapE in closed and open conformations,
determining a binding site of DapE or homologue thereof in the closed confirmation from the three-dimensional model, wherein DapE lacks a disulfide bond between Cys-155 and Cys-178,
separately determining the binding site of DapE or homologue thereof in the open confirmation, wherein DapE possess a disulfide bond between Cys-155 and Cys-178 and the lid domain is rotated by about 71° and
performing a computer fitting analysis to identify the candidate inhibitor that interacts with DapE in the open confirmation but does not interact with the binding site of DapE in the closed configuration, and prevents rotation of the DapE lid domain towards the DapE catalytic domain, wherein the interface of the catalytic domain and the lid domain comprises amino acid residues Glu-150, Ser-151, His-180, Gly-181, Glu-182, Lys-183, Arg-214, Asn-216, His-269, Arg-350, His-384, Tyr-385, Val-386, Asp-387, Lys-388, Thr-412, Gly-416, and His-440 of SEQ ID NO:2.
7. The method for identifying a candidate inhibitor of claim 6 , further comprising contacting the identified candidate inhibitor with DapE or homologue or active fragment thereof under conditions that allow for determination the effect of the inhibitor on S. aureus biological activity.
8. The method of claim 6 , wherein said structure coordinates of amino acids of DapE are as found in Table S1 and Table S2.