Variant, recombinant beta-glucocerebrosidase proteins with increased stability and increased retained catalytic activity
View Patent ↗Described herein are variant, recombinant β-glucocerebrosidase proteins characterized as having increased stability relative to recombinant wild-type β-glucocerebrosidase. Also provided herein are variant, recombinant β-glucocerebrosidase proteins characterized as retaining more catalytic activity relative to recombinant wild-type β-glucocerebrosidase. Further described herein are variant, recombinant β-glucocerebrosidase proteins that can have amino acid variations at one or more of the following positions: 316, 317, 321 and 145. Methods of making the variant, recombinant β-glucocerebrosidase proteins are also described as well as methods of treating patients having lysosomal storage diseases.
1. A variant, recombinant β-glucocerebrosidase protein comprising one of the following amino acid sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16.
2. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 2.
3. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 4.
4. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 5.
5. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 6.
6. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 7.
7. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 8.
8. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 9.
9. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 10.
10. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 11.
11. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 12.
12. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 13.
13. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 14.
14. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 15.
15. The protein of claim 1 , wherein the protein comprises SEQ ID NO: 16.
16. The protein of claim 1 , wherein the protein has increased stability relative to human wild-type, recombinant β-glucocerebrosidase.
17. The protein of claim 1 , wherein the protein retains more catalytic activity relative to human wild-type, recombinant β-glucocerebrosidase.
18. The protein of claim 17 , wherein more catalytic activity is retained at conditions of about neutral pH and about 37° C.
19. A method for treating a lysosomal storage disease, the method comprising administering to a human subject in need thereof the protein of claim 1 .
20. The method of claim 19 , wherein the lysosomal storage disease is Gaucher disease.