US2005176100A1PendingUtilityA1

Zace1: a human metalloenzyme

64
Priority: Nov 25, 1998Filed: Dec 8, 2004Published: Aug 11, 2005
Est. expiryNov 25, 2018(expired)· nominal 20-yr term from priority
Inventors:Paul O. Sheppard
C12Y 304/15001C07K 2319/00C12N 9/6489A61K 48/00A61K 38/00
64
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Claims

Abstract

Angiotensin-converting enzyme is a zinc metallopeptidase that plays roles in blood pressure regulation and fertility. The catalytic activities of angiotensin converting enzymes include the production of the potent vasopressor angiotensin H from angiotensin I, and the inactivation of the vasodilatory peptide bradykinin. Zace1 is a new form of human zinc metallopeptidase, which includes one zinc-dependent catalytic domain containing the motif “HEXXH” and one downstream “EX(I/V)X(D/S)” motif.

Claims

exact text as granted — not AI-modified
1 . An isolated polypeptide, comprising an amino acid sequence that is at least 70% identical to a reference amino acid sequence selected from the group consisting of: 
 (a) amino acid residues 367 to 430 of SEQ ID NO:1,    (b) amino acid residues 163 to 563 of SEQ ID NO:1,    (c) amino acid residues 52 to 563 of SEQ ID NO:1,    (d) amino acid residues 52 to 644 of SEQ ID NO:1,    (e) amino acid residues 52 to 648 of SEQ ID NO:1,    (f) amino acid residues 52 to 655 of SEQ ID NO:1,    (g) amino acid residues 52 to 662 of SEQ ID NO:1,    (h) amino acid residues 52 to 682 of SEQ ID NO:1,    (i) amino acid residues 52 to 694 of SEQ ID NO:1, and    (j) amino acid residues 1 to 694 of SEQ ID NO:1,    wherein the isolated polypeptide either (a) specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:1, or (b) exhibits dipeptidyl carboxypeptidase activity.    
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide has an amino acid sequence that is at least 80% identical to the reference amino acid sequence.  
     
     
         3 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide has an amino acid sequence that is at least 90% identical to the reference amino acid sequence.  
     
     
         4 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises an amino acid sequence selected from the group consisting of: (a) amino acid residues 367 to 430 of SEQ ID NO:1, (b) amino acid residues 163 to 563 of SEQ ID NO:1, (c) amino acid residues 52 to 563 of SEQ ID NO:1, (d) amino acid residues 52 to 644 of SEQ ID NO:1, (e) amino acid residues 52 to 648 of SEQ ID NO:1, (f) amino acid residues 52 to 655 of SEQ ID NO:1, (g) amino acid residues 52 to 662 of SEQ ID NO:1, (h) amino acid residues 52 to 682 of SEQ ID NO:1, (i) amino acid residues 52 to 694 of SEQ ID NO:1, and (j) amino acid residues 1 to 694 of SEQ ID NO:1.  
     
     
         5 . The isolated polypeptide of  claim 1 , wherein the polypeptide is a metallopeptidase.  
     
     
         6 . The isolated polypeptide of  claim 1 , wherein the polypeptide comprises an amino acid sequence comprising the motif “[GSTALIVN]-x-x-H-E-[LIVMFYW]-{DEHRKP}-H-x-[LIVMFYWGSPQ],” where “x” is any amino acid residue, acceptable amino acid residues are listed between square brackets, and unacceptable amino acid residues are listed between braces.  
     
     
         7 . The isolated polypeptide of  claim 6 , wherein the polypeptide comprises amino acid residues 395 to 404 of SEQ ID NO:1.  
     
     
         8 . A variant Zace1 polypeptide, wherein the amino acid sequence of the variant polypeptide shares an identity with the amino acid sequence of SEQ ID NO:1 selected from the group consisting of at least 70% identity, at least 80% identity, at least 90% identity, at least 95% identity, or greater than 95% identity, and wherein any difference between the amino acid sequence of the variant polypeptide and the amino acid sequence of SEQ ID NO:1 is due to one or more conservative amino acid substitutions.  
     
     
         9 . The isolated polypeptide of  claim 1 , comprising the amino acid sequence of SEQ ID NO:1.  
     
     
         10 . An isolated nucleic acid molecule that encodes polypeptide comprising an amino acid sequence selected from the group consisting of: (a) amino acid residues 367 to 430 of SEQ ID NO:1, (b) amino acid residues 163 to 563 of SEQ ID NO:1, (c) amino acid residues 52 to 563 of SEQ ID NO:1, (d) amino acid residues 52 to 644 of SEQ ID NO:1, (e) amino acid residues 52 to 648 of SEQ ID NO:1, (f) amino acid residues 52 to 655 of SEQ ID NO:1, (g) amino acid residues 52 to 662 of SEQ ID NO:1, (h) amino acid residues 52 to 682 of SEQ ID NO:1, (i) amino acid residues 52 to 694 of SEQ ID NO:1, and (j) amino acid residues 1 to 694 of SEQ ID NO:1.  
     
     
         11 . The isolated nucleic acid molecule of  claim 10 , wherein the nucleic acid molecule encodes amino acid residues 1 to 694 of SEQ ID NO:1.  
     
     
         12 . A vector, comprising the isolated nucleic acid molecule of  claim 11 .  
     
     
         13 . An expression vector, comprising the isolated nucleic acid molecule of  claim 11 , a transcription promoter, and a transcription terminator, wherein the promoter is operably linked with the nucleic acid molecule, and wherein the nucleic acid molecule is operably linked with the transcription terminator.  
     
     
         14 . A recombinant host cell comprising the expression vector of  claim 13 , wherein the host cell is selected from the group consisting of bacterium, yeast cell, fungal cell, insect cell, mammalian cell, and plant cell.  
     
     
         15 . A method of using the expression vector of  claim 13  to prepare Zace1 protein, comprising culturing recombinant host cells that comprise the expression vector and that produce the Zace1 protein.  
     
     
         16 . The method of  claim 15 , further comprising isolating the Zace1 protein from the cultured recombinant host cells.  
     
     
         17 . An antibody or antibody fragment that specifically binds with the polypeptide of  claim 4 .  
     
     
         18 . A method of detecting the presence of Zace1 in a biological sample, comprising: (a) contacting the biological sample with an antibody, or an antibody fragment, of  claim 17 , wherein the contacting is performed under conditions that allow the binding of the antibody or antibody fragment to the biological sample, and (b) detecting any of the bound antibody or bound antibody fragment.  
     
     
         19 . A fusion protein comprising a polypeptide of  claim 4 .  
     
     
         20 . An anti-idiotype antibody, or anti-idiotype antibody fragment, that specifically binds with the antibody or antibody fragment of  claim 13 , wherein the anti-idiotype antibody, or anti-idiotype antibody fragment, possesses dipeptidyl carboxypeptidase activity.  
     
     
         21 . The anti-idiotype antibody, or anti-idiotype antibody fragment of  claim 20 , wherein the anti-idiotype antibody, or anti-idiotype antibody fragment, possesses dipeptidyl carboxypeptidase activity.  
     
     
         22 . An isolated polypeptide comprising at least 15 contiguous amino acid residues of the amino acid sequence of SEQ ID NO:1.

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