Compositions of and methods of using sulfatases from
View Patent ↗This invention is related, in part, to sulfatase enzymes and methods of their use.
1. A method, comprising contacting a glycosaminoglycan with a glycosaminoglycan-degrading enzyme in an amount effective to cleave the glycosaminoglycan, thereby producing a cleavage product, wherein the glycosaminoglycan-degrading enzyme is an isolated sulfatase comprising the amino acid sequence of SEQ ID NO: 2 or 4.
2. The method of claim 1 , wherein the isolated sulfatase comprises the amino acid sequence of SEQ ID NO:2.
3. The method of claim 1 , wherein the isolated sulfatase comprises the amino acid sequence of SEQ ID NO:4.
4. The method of claim 1 , wherein the glycosaminoglycan is further contacted with at least one additional glycosaminoglycan-degrading enzyme.
5. The method of claim 4 , wherein the at least one additional glycosaminoglycan-degrading enzyme is used prior to, subsequent to, or concurrently with the sulfatase.
6. The method of claim 4 , wherein the at least one additional glycosaminoglycan-degrading enzyme is heparinase I, heparinse II, heparinase III, 2-O sulfatase, or Δ4,5 glycuronidase.
7. The method of claim 4 , wherein the at least one additional glycosaminoglycan-degrading enzyme is another sulfatase, glycosyl hydrolase, endoglucuronidase, or lyase.
8. The method of claim 6 , wherein the at least one additional glycosaminoglycan-degrading enzyme is Δ4,5 glycuronidase.
9. The method of claim 8 , wherein the Δ4,5 glycuronidase is contacted with the glycosaminoglycan prior to the sulfatase.
10. The method of claim 9 , wherein the glycosaminoglycan is further contacted with a 2-O sulfatase and the contact with the 2-O sulfatase is prior to the contact with the Δ4,5 glycuronidase.
11. The method of claim 10 , wherein the glycosaminoglycan is further contacted with a heparinase and the contact with the heparinase is prior to the contact with the 2-O sulfatase.
12. The method of claim 4 , wherein the sulfatase comprises the amino acid sequence of SEQ ID NO:2; wherein the glycosaminoglycan is also contacted with N-sulfamidase, and wherein the glycosaminoglycan is contacted with the sulfatase prior to the N-sulfamidase.
13. The method of claim 11 , wherein the sulfatase comprises the amino acid sequence of SEQ ID NO:2; wherein the glycosaminoglycan is also contacted with N-sulfamidase; and wherein the glycosaminoglycan is contacted with the sulfatase prior to the N-sulfamidase.
14. The method of claim 1 , wherein the glycosaminoglycan is 3-O desulfated prior to contact with the sulfatase.
15. The method of claim 1 , further comprising analyzing the cleavage product.
16. The method of claim 15 , wherein the glycosaminoglycan is also contacted with at least one additional glycosaminoglycan-degrading enzyme.
17. The method of claim 16 , wherein the at least one additional glycosaminoglycan-degrading enzyme is used prior to, subsequent to, or concurrently with the sulfatase.
18. The method of claim 16 , wherein the at least one additional glycosaminoglycan-degrading enzyme is heparinase I, heparinse II, heparinase III, 2-O sulfatase, or Δ4,5 glycuronidase.
19. The method of claim 16 , wherein the at least one additional glycosaminoglycan-degrading enzyme is another sulfatase, glycosyl hydrolase, endoglucuronidase, or lyase.
20. The method of claim 18 , wherein the at least one additional glycosaminoglycan-degrading enzyme is Δ4,5 glycuronidase.
21. The method of claim 20 , wherein the Δ4,5 glycuronidase is contacted with the glycosaminoglycan prior to the sulfatase.
22. The method of claim 21 , wherein the glycosaminoglycan is further contacted with a 2-O sulfatase and the contact with the 2-O sulfatase is prior to the contact with the Δ4,5 glycuronidase.
23. The method of claim 22 , wherein the glycosaminoglycan is further contacted with a heparinase and the contact with the heparinase is prior to the contact with the 2-O sulfatase.
24. The method of claim 16 , wherein the sulfatase comprises the amino acid sequence of SEQ ID NO:2; wherein the glycosaminoglycan is also contacted with an N-sulfamidase; and wherein the glycosaminoglycan is contacted with the sulfatase prior to the N-sulfamidase.
25. The method of claim 23 , wherein the sulfatase comprises the amino acid sequence of SEQ ID NO:2; wherein the glycosaminoglycan is also contacted with an N-sulfamidase; and wherein the glycosaminoglycan is contacted with the sulfatase prior to the N-sulfamidase.
26. The method of claim 15 , wherein the glycosaminoglycan is 3-O desulfated prior to contact with the sulfatase.
27. The method of claim 15 , wherein the presence of a particular glycosaminoglycan in a sample is determined based on the result from the analyzing step.
28. The method of claim 15 , wherein the purity of a glycosaminoglycan in a sample is determined based on the result from the analyzing step.
29. The method of claim 15 , wherein the composition of a glycosaminoglycan in a sample is determined based on the result from the analyzing step.
30. The method of claim 15 , wherein the sequence of saccharide units in a glycosaminoglycan is determined based on the result from the analyzing step.
31. The method of claim 15 , wherein the analyzing step is performed by subjecting the cleavage product to mass spectrometry, NMR spectroscopy, gel electrophoresis, capillary electrophoresis, or HPLC.