PAT 02-25-03 06524807 NDN- 217-0467-0451-1

INVENTOR(S)- Talanian, Robert V.; Mankovich, John A.; Ghayur, Tariq; Ferenz, Catherine R.

PATENT NUMBER- 06524807
PATENT APPLICATION NUMBER- 827708
DATE FILED- 2001-04-06
PATENT DATE- 2003-02-25
NUMBER OF CLAIMS- 9
EXEMPLARY CLAIMS- 1
FIGURES- 1
ART/GROUP UNIT- 1652
PATENT CLASS- Invention (utility) patent
PATENT ASSIGNEE(S)- BASF Aktiengellschaft
ASSIGNEE CITY- Rheinland-Pfalz
ASSIGNEE COUNTRY- DEX
ATTORNEY, AGENT, OR FIRM- Lahive & Cockfield, LLP; DeConti, Jr., Giulio A.; Soroos, Cynthia M.
U.S. PATENT CLASS- 435023000O
U.S. CLASSIFICATION REFS.- X435219000; X435226000; X435252300; X435320100; X536023200; X536023500
INTERNATIONAL PATENT CLASS- 7C12Q00137; C12N00950; C12N00120; C12N01500; C07H02104
PATENT REFERENCE(S)- 5416013; 5492824; 5869315
FOREIGN DOCUMENT REFERENCE (S)- WO 91/15577; WO 93/05071; WO 94/00154; WO 95/00160
FOREIGN COUNTRY CODE- WOX; WOX; WOX; WOX

Modified forms of human interleukin-1(small beta, Greek) converting enzyme (ICE) that display proteolytic activity and, furthermore, have increased stability compared to unmodified human ICE are disclosed. Nucleic acid molecules encoding a modified p10 subunit of ICE, and recombinant vectors and host cells incorporating such nucleic acid molecules, are also disclosed. A modified ICE protein of the invention can be used to cleave proteolytically ICE substrates and to identify modulators of ICE activity in screening assays. Moreover, due to its enhanced stability, the modified ICE of the invention is particularly suitable for use in the preparation of ICE crystals for X-ray crystallography.

EXEMPLARY CLAIMS- 1. A method for cleaving an interleukin-1(small beta, Greek) converting enzyme (ICE) substrate, comprising contacting the substrate with a modified ICE such that the ICE substrate is cleaved, wherein the modified ICE comprises an amino acid sequence having an amino acid corresponding to aspartic acid at position 381 of unmodified ICE (SEQ ID NO: 2) replaced with a mutant amino acid structure, the mutant amino acid structure being capable of forming a salt bridge with an amino acid corresponding to arginine at position 383 of unmodified ICE.