United States Patent Application 20020052474
Kind Code A1
Shu, Hong-Bing ; et al. May 2, 2002
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Regulators of apoptosis
Abstract
The invention provides methods and compositions relating to apopotosis regulating proteins, known as Casper proteins, and related nucleic acids. The proteins may be produced recombinantly from transformed host cells from the disclosed Casper encoding nucleic acid or purified from human cells. The invention provides specific hybridization probes and primers capable of specifically hybridizing with the disclosed Casper gene, Casper-specific binding agents such as specific antibodies, and methods of making and using the subject compositions in diagnosis, therapy and in the biopharmaceutical industry.
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Inventors: Shu, Hong-Bing; (South San Francisco, CA) ; Goeddel, David V.; (Hillsborough, CA)
Correspondence Name and Address: RICHARD ARON OSMAN
SCIENCE AND TECHNOLOGY LAW GROUP
75 DENISE DRIVE
HILLSBOROUGH
CA
94010
Serial No.: 861270
Series Code: 09
Filed: May 18, 2001
U.S. Current Class: 530/350; 435/226; 435/325; 435/69.1; 536/23.5
U.S. Class at Publication: 530/350; 435/226; 435/325; 435/69.1; 536/23.5
Intern'l Class: C12N 009/64; C12P 021/02; C07H 021/04; C12N 005/06
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Claims
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1. An isolated Casper protein comprising SEQ ID NO: 2 or a fragment thereof having Casper-specific activity.
2. An isolated protein according to claim 1, wherein said protein specifically binds at least one of a FADD, TRAF1, TRAF2, Caspase-3 or Caspase-8 protein.
3. An isolated protein according to claim 1, wherein said protein comprises a deletion mutant of SEQ ID NO:2, said deletion mutant comprising SEQ ID NO:2, residues 1-96, 1-202, 1-435, 78-480, 192-435, 192-480 or 390-480.
4. An isolated protein according to claim 1, wherein said protein comprises SEQ ID NO:2, residue 360 (tyrosine 360) joined directly to at least 6 residues of SEQ ID NO:2 flanking said tyrosine 360.
5. An isolated protein according to claim 1, wherein said protein comprises SEQ ID NO:2, residue 360 (tyrosine 360) joined directly to at least six residues of SEQ ID NO:2 flanking said tyrosine 360, wherein three of said six residues are located on each side of said tyrosine 360.
6. A recombinant nucleic acid encoding a protein according to claim 1.
7. A cell comprising a nucleic acid according to claim 3.
8. A method of making an isolated Casper protein, comprising steps: introducing a nucleic acid according to claim 3 into a host cell or cellular extract, incubating said host cell or extract under conditions whereby said nucleic acid is expressed as a transcript and said transcript is expressed as a translation product comprising said protein, and isolating said translation product.
6. An isolated Casper protein made by the method of claim 5.
9. An isolated Casper nucleic acid comprising SEQ ID NO: 1, or a fragment thereof having at least 24 consecutive bases of SEQ ID NO: 1 and sufficient to specifically hybridize with a nucleic acid having the sequence of SEQ ID NO: 1.
10. A method of screening for an agent which modulates the binding of a Casper protein to a binding target, said method comprising the steps of: incubating a mixture comprising: an isolated protein according to claim 1, a binding target of said protein, and a candidate agent; under conditions whereby, but for the presence of said agent, said protein specifically binds said binding target at a reference affinity; detecting the binding affinity of said protein to said binding target to determine an agent-biased affinity, wherein a difference between the agent-biased affinity and the reference affinity indicates that said agent modulates the binding of said protein to said binding target.
11. A method according to claim 10, wherein said binding target is a one of a FADD, TRAF1, TRAF2, Caspase-3 or Caspase-8 protein. |
