Modulation of COP9 signalsome isopeptidase activity
AssigneeCalifornia Institute of Technology
InventorsRaymond J. Deshaies, Gregory Cope, Rati Verma, Xavier I. Ambroggio
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Abstract
Methods for identifying agents that can increase or decrease the isopeptidase activity of a COP9 signalsome (CSN) are provided, as are agent identified using such screening assays. In addition, methods of ameliorating a pathologic condition such as a cancer or an autoimmune disease in a subject by modulating the CSN isopeptidase activity are provided, as are medicaments useful for treating a subject having such a condition.
Claims86
An isolated polypeptide comprising a mutant JAMM domain, said JAMM domain comprising an amino acid sequence of HXHXXXXXXXXXXD (SEQ ID NO:
- 1),
- wherein H is histidine, D is aspartate, and X is any amino acid, and
- wherein, in said mutant JAMM domain, at least one of said H residues is substituted with any amino acid residue other than H, said D residue is substituted with an amino acid residue other than D, or a combination thereof.
An isolated polypeptide comprising a mutant JAMM domain, said JAMM domain comprising an amino acid sequence of H(S/T)HXXXXXXXSXXD (SEQ ID NO:
- 65),
- wherein H is histidine, D is aspartate, S is serine, (S/T) is serine or threonine, and X is any amino acid, and
- wherein, in said mutant JAMM domain, at least one of said H residues is substituted with an amino acid residue other than H;
- said D residue is substituted with an amino acid residue other than D;
- said S residue is substituted with an amino acid residue other than S;
- said T residue, when present, is substituted with an amino acid residue other than T, or a combination thereof.
An isolated polypeptide comprising a mutant JAMM domain, said JAMM domain comprises an amino acid sequence of EXHyXHyHy(X)nHXHXXXXXXXXXXD (SEQ ID NO:
- 13),
- wherein H is histidine, D is aspartate, E is glutamic acid, Hy is a hydrophobic amino acid residue, X is any amino acid, and (X)n indicates about 40 to 83 amino acid residues, and
- wherein, in said mutant JAMM domain, at least one of said H residues is substituted with an amino acid residue other than H;
- said D residue is substituted with an amino acid residue other than D;
- said E residue is substituted with an amino acid residue other than E;
- or a combination thereof.
A method of identifying an inhibitor of an isopeptidase activity of a polypeptide by rational drug design, wherein the polypeptide comprises a JAMM domain consisting essentially of an amino acid sequence of HXHXXXXXXXXXXD (SEQ ID NO:
- 1), and wherein H is histidine, D is aspartate, and X is any amino acid, the method comprising:
- designing a potential inhibitor for the polypeptide that will form a bond with the JAMM domain based upon the crystal structure co-ordinates of the polypeptide, synthesizing the potential inhibitor, and
- detecting inhibition of isopeptidase activity of the polypeptide by the potential inhibitor, thereby identifying an inhibitor of the polypeptide.
A method of identifying an inhibitor of an isopeptidase activity of a polypeptide by rational drug design, wherein the polypeptide comprises a JAMM domain consisting essentially of an amino acid sequence of H(S/T)HXXXXXXXSXXD (SEQ ID NO:
- 65), wherein H is histidine, D is aspartate, S is serine, (S/T) is serine or threonine, and X is any amino acid, the method comprising:
- designing a potential inhibitor for the polypeptide that will form a bond with the JAMM domain based upon the crystal structure co-ordinates of the polypeptide, synthesizing the potential inhibitor, and
- detecting inhibition of isopeptidase activity of the polypeptide by the potential inhibitor, thereby identifying an inhibitor of the polypeptide.
A method of identifying an agent that modulates an isopeptidase activity of a polypeptide comprising a JAMM domain, said JAMM domain comprising an amino acid sequence of HXHXXXXXXXXXXD (SEQ ID NO:
- 1), and wherein H is histidine, D is aspartate, and X is any amino acid, the method comprising:
- contacting the polypeptide comprising the JAMM domain and a test agent under conditions suitable for isopeptidase activity, wherein said isopeptidase activity cleaves a modifier protein from a target protein, and
- detecting a change in isopeptidase activity by the polypeptide in the presence of the test agent as compared to the absence of the test agent, thereby identifying an agent that modulates isopeptidase activity of the polypeptide comprising the JAMM domain.
A method of treating a condition by modulating protein degradation in a cell associated with the condition, comprising administering to the subject at least a first agent that modulates an isopeptidase activity of a polypeptide comprising a JAMM domain, said JAMM domain having an amino acid sequence of HXHXXXXXXXXXXD (SEQ ID NO:
- 1), and wherein H is histidine, D is aspartate, and X is any amino acid.
An isolated JAMM domain, consisting essentially of a JAMM domain having an amino acid sequence of EXHyXHyHy(X)nHXHXXXXXXXXXXD (SEQ ID NO:
- 13), wherein E is glutamic acid, Hy is a hydrophobic amino acid residue, X is any amino acid, and (X)nindicates about 40 to 83 amino acid residues.
Dependencies are inferred from the claim text.