Inhibitor of NF-KB activation
View Patent ↗The present invention relates to a protein, designated NAP (Nemo Associated Protein), that was discovered to inhibit the activation of transcription factor NF-kB by various signals that are important in inflammatory and immune processes. The DNA and the recombinant production of this protein is also disclosed as well as methods of using the protein or encoding DNA.
1. An isolated protein which is capable of binding to tumor necrosis factor receptor-associated 2 protein (TRAF2), said protein comprising:
(A) a polypeptide of SEQ ID NO:3; or
(B) a variant that has no more than ten amino acid changes from the amino acid sequence of SEQ ID NO:3, wherein said variant is capable of binding to TRAF2.
2. The isolated protein of claim 1 , which is a protein comprising the amino acid sequence of SEQ ID NO:3.
3. A composition comprising the isolated protein of claim 1 and a pharmaceutically acceptable excipient, diluent, or auxiliary agent.
4. A molecule having the binding portion of an antibody capable of binding to the portion of said isolated protein of claim 1 that is said polypeptide of (A) or said variant of (B).
5. The molecule of claim 4 , which is an antibody.
6. The molecule of claim 5 , wherein said antibody is a monoclonal antibody.
7. A composition comprising the molecule of claim 4 , and a pharmaceutically acceptable excipient, diluent, or auxiliary agent.
8. An isolated protein in accordance with claim 1 , wherein said protein and said variant are each capable of binding to a component of the NF-κB complex selected from the group consisting of IKappaB kinase complex associated protein (IKAP), IKappaB kinase-alpha (IKK-alpha), IKappaB kinase-beta (IKK-beta), IKappaB kinase-gamma (IKK-gamma) and NF-κB inducing kinase (NIK).
9. An isolated protein in accordance with claim 1 , comprising a variant of the polypeptide of SEQ ID NO:3, which variant has no more than ten amino acid changes from the amino acid sequence of SEQ ID NO:3, and which variant is capable of binding to TRAF2.
10. The isolated protein of claim 1 , wherein said variant of (B) has no more than five amino acid changes from the amino acid sequence of SEQ ID NO:3.
11. The isolated protein of claim 1 , wherein each said change from the amino acid sequence of SEQ ID NO:3 is a conservative substitution selected from among the substitutions in the following list:
Original
Residue
Substitution
Ala
Gly; Ser
Arg
Lys
Asn
Gln; His
Asp
Glu
Cys
Ser
Gln
Asn
Glu
Asp
Gly
Ala; Pro
His
Asn; Gln
Ile
Leu; Val
Leu
Ile; Val
Lys
Arg; Gln; Glu
Met
Leu; Tyr; Ile
Phe
Met; Leu; Tyr
Ser
Thr
Thr
Ser
Trp
Tyr
Tyr
Trp; Phe
Val
Ile; Leu
or a conservative substitution that is an exchange within one of the following five groups:
Small aliphatic, nonpolar or
slightly polar residues:
Ala, Ser, Thr, Pro, Gly;
Polar negatively charged
Asp, Asn, Glu, Gln;
residues and their amides:
Polar, positively oharged residues:
His, Arg, Lys;
Large aliphatic nonpolar residues:
Met, Leu, Ile, Val, Cys;
and
Large aromatic residues:
Phe, Tyr, Trp.
12. The isolated protein of claim 11 , wherein said variant has no more than 5 of said amino acid changes from the amino acid sequence of SEQ ID NO:3.
13. A molecule having the binding portion of an antibody capable of binding to the polypeptide of SEQ ID NO:3.
14. The molecule of claim 13 , which is an antibody.
15. The molecule of claim 14 , wherein said antibody is a monoclonal antibody.
16. An isolated protein comprising:
A) a polypeptide of SEQ ID NO: 3; or
B) a variant that has no more than 10 amino acid changes from the amino acid sequence of SEQ ID NO:3.
17. The isolated protein of claim 16 , wherein said variant has no more than 5 of said amino acid changes from the amino acid sequence of SEQ ID NO:3.