Glycosylation and functionality of recombinant β-glucocerebrosidase from various production systems
| Autor: | Tali Kizhner, Myriam Golembo, David Aviezer, Anna Gantman, Mariana Hainrichson, Salit Tzaban, Yoseph Shaaltiel, Yoram Tekoah |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Male
β-glucocerebrosidase Glycosylation Mannose Gaucher disease Biochemistry 2AB 2-aminobenzamide ICR imprinting control region law.invention Mice chemistry.chemical_compound law Enzyme Stability Carbohydrate Conformation MALDI–TOF-MS matrix-assisted laser desorption ionization–time-of-flight-MS Tissue Distribution chemistry.chemical_classification Mice Inbred ICR GU glucose unit GD Gaucher disease PNGase F peptide N-glycosidase F Recombinant Proteins Taliglucerase alfa ERT enzyme replacement therapy Carbohydrate Sequence lysosomal storage disease Recombinant DNA Glucosylceramidase ERT medicine.drug HSD honestly significant difference SA sialic acid Imiglucerase Molecular Sequence Data Biophysics PNGase A peptide N-glycosidase A macrophage Biology PNP-G p-nitrophenyl β-D-glucopyranoside S2 Cell Line 4MU-G 4-methylumbelliferyl β-D-glucopyranoside Macrophages Alveolar NP normal phase medicine Animals Humans Molecular Biology MBL mannose-binding lectin Original Paper Velaglucerase alfa Biological Transport Cell Biology MR mannose receptor Rats Kinetics Enzyme chemistry CHO Chinese-hamster ovary Protein Processing Post-Translational Glucocerebrosidase DAPI 4′ 6-diamidino-2-phenylindole |
| Zdroj: | Bioscience Reports |
| ISSN: | 1573-4935 0144-8463 |
| DOI: | 10.1042/bsr20130081 |
| Popis: | The glycosylation of recombinant β-glucocerebrosidase, and in particular the exposure of mannose residues, has been shown to be a key factor in the success of ERT (enzyme replacement therapy) for the treatment of GD (Gaucher disease). Macrophages, the target cells in GD, internalize β-glucocerebrosidase through MRs (mannose receptors). Three enzymes are commercially available for the treatment of GD by ERT. Taliglucerase alfa, imiglucerase and velaglucerase alfa are each produced in different cell systems and undergo various post-translational or post-production glycosylation modifications to expose their mannose residues. This is the first study in which the glycosylation profiles of the three enzymes are compared, using the same methodology and the effect on functionality and cellular uptake is evaluated. While the major differences in glycosylation profiles reside in the variation of terminal residues and mannose chain length, the enzymatic activity and stability are not affected by these differences. Furthermore, the cellular uptake and in-cell stability in rat and human macrophages are similar. Finally, in vivo studies to evaluate the uptake into target organs also show similar results for all three enzymes. These results indicate that the variations of glycosylation between the three regulatory-approved β-glucocerebrosidase enzymes have no effect on their function or distribution. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|