Nucleic acids encoding secreted polypeptides that stimulate release of proteoglycans from cartilage
View Patent ↗The present invention is directed to secreted and transmembrane polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.
1. An isolated nucleic acid encoding a polypeptide which stimulates release of proteoglycans from cartilage tissue and having at least 95% nucleic acid sequence identity to:
(a) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2);
(b) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2), lacking its associated signal peptide;
(c) the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1);
(d) the full-length coding sequence of the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1); or
(e) the full-length coding sequence of the cDNA deposited under ATCC accession number 209526.
2. The isolated nucleic acid of claim 1 , encoding a polypeptide which stimulates release of proteoglycans from cartilage tissue and having at least 99% nucleic acid sequence identity to:
(a) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2);
(b) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2), lacking its associated signal peptide;
(c) the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1);
(d) the full-length coding sequence of the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1); or
(e) the full-length coding sequence of the cDNA deposited under ATCC accession number 209526.
3. An isolated nucleic acid comprising:
(a) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2);
(b) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2), lacking its associated signal peptide;
(c) the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1);
(d) the full-length coding sequence of the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1); or
(e) the full-length coding sequence of the cDNA deposited under ATCC accession number 209526.
4. An isolated nucleic acid comprising a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2).
5. An isolated nucleic acid comprising a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2), lacking its associated signal peptide.
6. An isolated nucleic acid comprising the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1).
7. An isolated nucleic acid comprising the full-length coding sequence of the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1).
8. An isolated nucleic acid comprising the full-length coding sequence of the cDNA deposited under ATCC accession number 209526.
9. An isolated nucleic acid that hybridizes under high stringency conditions to:
(a) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2);
(b) a nucleic acid sequence encoding the polypeptide shown in FIG. 2 (SEQ ID NO:2), lacking its associated signal peptide;
(c) the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1);
(d) the full-length coding sequence of the nucleic acid sequence shown in FIG. 1 (SEQ ID NO:1); or
(e) the full-length coding sequence of the cDNA deposited under ATCC accession number 209526
wherein said isolated nucleic acid encodes a polypeptide which stimulates release of proteoglycans from cartilage tissue and wherein said high stringency conditions are selected from the group consisting of: (i) 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50° C.; (ii) 50% (v/v) formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6/5 with 750 mM sodium chloride, 75 mM sodium citrate at 42° C.; and (iii) 50% formamide, 5×SSC (0.75 M sodium chloride, 0.075 M Sodium citrate), 50 mM sodium phosphate (pH 6.8),0.1% sodium pyrophosphhate, 5× Denhardt's solution, sonicated salmon sperm DNA (50 μg/ml), 0.1% sodium dodecyl sulphate, and 10% dextran sulfate at 42° C., with washes at 42° C. in 0.2×SSC (0.75 M sodium chloride, 0.075 M sodium citrate) and 50% formamide at 55° C. followed by a high-stringency wash consisting of 0.1×SSC (0.75 M sodium chloride, 0.075 N sodium citrate) containing EDTA at 55° C.
10. An isolated vector comprising the nucleic acid of claim 1 .
11. The isolated vector of claim 10 , wherein said nucleic acid is operably linked to control sequences recognized by a host cell transformed with the vector.
12. An isolated host cell comprising the vector of claim 10 .
13. The isolated host cell of claim 12 , wherein said cell is a CHO cell, an E. coli or a yeast cell.