2-O sulfatase nucleic acid compositions
The invention relates to 2-O sulfatase and uses thereof. In particular, the invention relates to recombinantly produced 2-O sulfatase, functional variants and nucleic acid molecules that encode these molecules. The invention also provides methods of using 2-O sulfatase for a variety of purposes, including degrading and analyzing glycosaminoglycans (GAGs) present in a sample. For instance, 2-O sulfatase may be used for determining the purity, identity, composition and sequence of glycosaminoglycans present in a sample. The invention also relates to methods of inhibiting angiogenesis and cellular proliferation as well as methods for treating cancer, neurodegenerative disease, atherosclerosis and microbial infection using 2-O sulfatase and/or GAG fragments produced by degradation with 2-O sulfatase.
1. An isolated nucleic acid molecule selected from the group consisting of:
(a) nucleic acid molecules which hybridize under stringent conditions to a nucleic acid molecule having a nucleotide sequence selected from the group consisting of nucleotide sequences set forth as SEQ ID NOs: 1 and 3, and which code for a 2-O sulfatase,
wherein the hybridization conditions are 1) hybridization at 65° C. in hybridization buffer that consists of 3.5×SSC, 0.02% Ficoll, 0.02% polyvinyl pyrrolidone, 0.02% Bovine Serum Albumin, 2.5 mM NaH2PO4, pH 7, 0.5% sodium dodecyl sulphate (SDS), 2 mM ethylenediaminetetracetic acid (EDTA), wherein SSC is 0.15M sodium chloride/0.015M sodium citrate, pH 7 and 2) washing in 2×SSC at room temperature and then in 0.1×SSC/0.1×SDS at 68° C.,
(b) nucleic acid molecules that differ from the nucleic acid molecules of (a) in codon sequence due to degeneracy of the genetic code, and
(c) full complements of (a) or (b).
2. The isolated nucleic acid molecule of claim 1 , wherein the isolated nucleic acid molecule comprises a nucleic acid sequence set forth as SEQ ID NO: 3.
3. The isolated nucleic acid molecule of claim 1 , wherein the isolated nucleic acid molecule codes for SEQ ID NO: 4.
4. An isolated nucleic acid molecule comprising a nucleotide sequence that is at least 90% identical to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1 and 3, which codes for a 2-O sulfatase that has an activity of the 2-O sulfatase set forth as SEQ ID NO: 2 or 4.
5. The isolated nucleic acid molecule of claim 4 , wherein the nucleotide sequence is at least 95% identical.
6. The isolated nucleic acid molecule of claim 5 , wherein the nucleotide sequence is at least 97% identical.
7. The isolated nucleic acid molecule of claim 6 , wherein the nucleotide sequence is at least 98% identical.
8. The isolated nucleic acid molecule of claim 7 , wherein the nucleotide sequence is at least 99% identical.
9. The isolated nucleic acid molecule of claim 8 , wherein the nucleotide sequence is at least 99.5% identical.
10. The isolated nucleic acid molecule of claim 9 , wherein the nucleotide sequence is at least 99.9% identical.
11. An expression vector comprising the isolated nucleic acid molecule of claim 1 operably linked to a promoter.
12. An expression vector comprising the isolated nucleic acid molecule of claim 4 operably linked to a promoter.
13. A host cell comprising the expression vector of claim 11 .
14. A host cell comprising the expression vector of claim 12 .
15. A composition comprising the nucleic acid of claim 1 and a pharmaceutically acceptable carrier.
16. A composition comprising the nucleic acid of claim 4 and a pharmaceutically acceptable carrier.
17. A composition comprising:
the expression vector of claim 11 and a pharmaceutically acceptable carrier.
18. A composition comprising:
the expression vector of claim 12 and a pharmaceutically acceptable carrier.
19. A composition comprising:
the host cell of claim 13 and a pharmaceutically acceptable carrier.
20. A composition comprising:
the host cell of claim 14 and a pharmaceutically acceptable carrier.