Claims
- 1. An isolated protein, consisting of a protein product of a gene which is structurally related to the ced-3 and ICE genes, said isolated protein
having an alteration in the amino acid sequence of the product of a gene which is structurally related to the Ced-3 and ICE genes, said alteration corresponding to an alteration in the sequence of SEQ ID NO: 4 selected from the group consisting of:
i) L to F at amino acid 26; ii) G to R at amino acid 65; iv) G to S at amino acid 287; v) truncation of said protein after amino acid 323; vi) truncation of said protein after amino acid 339; vii) A to V at amino acid 361; viii) E to K at amino acid 390; and ix) T to F at amino acid 393.
- 2. The protease of claim 1 which cleaves after aspartate residues.
- 3. The protease of claim 1 which is a cysteine protease.
- 4. An isolated ICE polypeptide (SEQ ID NO: 4) having an alteration which reduces the activity of the enzyme, wherein said alteration is selected from the group consisting of:
a) L to Fat amino acid 26; b) G to R at amino acid 65; c) G to S at amino acid 287; d) truncation of said polypeptide after amino acid 323; e) truncation of said polypeptide after amino acid 339; f) A to V at amino acid 361; g) E to K at amino acid 390; and h) T to F at amino acid 393.
- 5. A product of the gene of claim 4 selected from RNA and protein.
- 6. A constitutively activated cell death protein comprising an amino acid sequence, said sequence comprising a portion of the Ced-3 protein shown in SEQ ID NO: 2 of FIG. 6A, said portion selected from the group consisting of:
a) the amino acids from approximately 150 to 503 (SEQ ID NO: 20); b) the amino acids from approximately 373 to 503 (SEQ ID NO: 21); and c) the amino acids from approximately 150 to 372 (SEQ ID NO: 22).
- 7. The constitutively activated cell death protein of claim 6, further comprising a subportion of the region of Ced-3 from amino acids 1 to 149, as shown in SEQ ID NO: 2 of FIG. 6A, said subportion enhancing the activity of the protein.
- 8. A constitutively activated cell death protein having an amino acid sequence ICE from the sequence shown in FIG. 6A (SEQ ID NO: 4), said sequence selected from the group consisting of:
a) the amino acids from approximately 111 to 404 (SEQ ID NO: 23); b) the amino acids from approximately 298 to 404 (SEQ ID NO: 24); c) the amino acids from approximately 111 to 297 (SEQ ID NO: 25).
- 9. An isolated protein which is the NEDD-2 protein (SEQ ID NO: 26) having an alteration which inactivates the protein, wherein said alteration is A to V at amino acid 117.
- 10. The isolated protein of claim 9, wherein said alteration is C to A at amino acid 303.
- 11. The isolated protein of claim 9, wherein said alteration is C to S at amino acid 303.
- 12. Isolated protein which is selected from the group consisting of Ced-3 (SEQ ID NO: 2), ICE (SEQ ID NO: 4), and NEDD-2 (SEQ ID NO: 13), said protein having an alteration at a conserved amino acid corresponding to an amino acid of the Ced-3 protein (SEQ ID NO. 2) selected from the group consisting of:
a) Ced-3 Ser 183 or ICE Ser 126; b) Ced-3 Met 234; c) Ced-3 Arg 242; d) Ced-3 Leu 246 or ICE Leu 166; e) Ced-3 Ile 247 or ICE Ile 167; f) Ced-3 Ile 248 or ICE Ile 168; g) Ced-3 Asn 250 or ICE Asn 170; h) Ced-3 Phe 253 or ICE Phe 173; i) Ced-3 Arg 259 or ICE Arg 179; j) Ced-3 Gly 261 or ICE Gly 181; k) Ced-3 Asp 265 or ICE Asp 185; l) Ced-3 Gly 277 or ICE Gly 197; m) Ced-3 Tyr 278 or ICE Tyr 198; n) Ced-3 Val 280 or ICE Val 200; o) Ced-3 Lys 283 or ICE Lys 203; p) Ced-3 Asn 285 or ICE Asn 205; q) Ced-3 Leu 286 or ICE Leu 206; r) Ced-3 Thr 287 or ICE Thr 207; s) Ced-3 Met 291 or ICE Met 211; t) Ced-3 Phe 298 or ICE Phe 218; u) Ced-3 His 304 or ICE His 224; v) Ced-3 Asp 306 or ICE Asp 228; w) Ced-3 Ser 307, ICE Ser 229, or NEDD-2 Ser 16; x) Ced-3 Leu 310, ICE Leu 232, or NEDD-2 Val 19; y) Ced-3 Val 311, or ICE Val 233, or NEDD-2 Val 20; z) Ced-3 Ser 314 or ICE Ser 236; aa) Ced-3 His 315 or ICE His 237; bb) Ced-3 Gly 316 or ICE Gly 238; cc) Ced-3 Ile 321, ICE Ile 243, or NEDD-2 Leu 23; dd) Ced-3 Gly 323, ICE Gly 245, or NEDD-2 Asp 25; ee) Ced-3 le 334, ICE Ile 261, or NEDD-2 Phe 31; ff) Ced-3 Asn 339 or ICE Asn 266; gg) Ced-3 Pro 344 or ICE Pro 271; hh) Ced-3 Leu 346 or ICE Leu 273; ii) Ced-3 Lys 349 or ICE Lys 276; jj) Ced-3 Pro 350, ICE Pro 277, or NEDD-2 Pro 37; kk) Ced-3 Lys 351 or ICE Lys 278; ll) Ced-3 Gln 356, ICE Gln 283, or NEDD-2 Glu 43; mm) Ced-3 Ala 357, ICE Ala 284, or NEDD-2 Thr 44; nn) Ced-3 Cys 358 or ICE Cys 285; oo) Ced-3 Arg 359, ICE Arg 286 or NEDD-2 Arg 46; pp) Ced-3 Gly 360, ICE Gly 287, or NEDD-2 Gly 47; qq) Ced-3 Asp 371 or ICE Asp 297; rr) Ced-3 Asp 414, ICE Asp 326, or NEDD-2 Asp 82; ss) Ced-3 Arg 429, ICE Arg 341, or NEDD-2 Arg 97; tt) Ced-3 Gly 434, ICE Gly 346, or NEDD-2 Gly 102; uu) Ced-3 Ser 435, ICE Ser 347, or NEDD-2 Ser 103; vv) Ced-3 Ile 438, ICE Ile 350; NEDD-2 Ile 106; ww) Ced-3 Ala 449, ICE Ala 361, or NEDD-2 Ala 108; xx) Ced-3 Val 454, ICE Val 366, or NEDD-2 Val 123; yy) Ced-3 Leu 488, ICE Leu 394, or NEDD-2 Leu 158; zz) Ced-3 Tyr 493, ICE Tyr 399, or NEDD-2 Tyr 163; and aaa) Ced-3 Pro 496, ICE Pro 402, or NEDD-2 Pro 166.
- 13. The isolated protein selected from the group consisting of Ced-3 (SEQ ID NO: 2) and ICE (SEQ ID NO: 4), said protein having an amino acid alteration in an amino acid corresponding to Cys 358 of Ced-3, and Cys 285 of ICE.
- 14. The isolated protein of claim 13, wherein said alteration is a Cys to Ala alteration.
- 15. The isolated protein of claim 13, wherein said protein is ICE and said alteration is at conserved amino acid 285 of said ICE.
- 16. The isolated protein of claim 13, wherein said protein is NEDD-2 and said alteration is at conserved amino acid 303 of said NEDD-2.
- 17. Isolated nucleic acid encoding the protein of claim 12.
- 18. A method of preventing cell death, said method comprising administering a polypeptide of claim 12.
- 19. The method of claim 14, wherein said administering is to a patient and said polypeptide is provided at a therapeutically effective dose.
- 20. A method of preventing cell death, said method comprising administering a therapeutically effective amount of the isolated nucleic acid of claim 17.
RELATED APPLICATION
[0001] This application is a continuation of U.S. Ser. No. 08/394,189, filed Feb. 24, 1995, which is a continuation-in-part of U.S. Ser. No. 08/282,211, filed Jul. 11, 1994; which is a divisional of Ser. No. 07/984,182, filed Nov. 20, 1992, now abandoned; which is a continuation-in-part of U.S. Ser. No. 07/897,788, filed Jun. 12, 1992, now abandoned. The teachings of U.S. Ser. No. 07/897,788 are incorporated by reference.
GOVERNMENT FUNDING
[0002] Work described herein was supported by grants GM24663 and GM24943 from the U.S. Public Health Service. The U.S. Government has certain rights in the invention.
Divisions (1)
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07984182 |
Nov 1992 |
US |
Child |
08282211 |
Jul 1994 |
US |
Continuations (2)
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09083662 |
May 1998 |
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09888243 |
Jun 2001 |
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08394189 |
Feb 1995 |
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09083662 |
May 1998 |
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Continuation in Parts (2)
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07897788 |
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07984182 |
Nov 1992 |
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