Download MendeleyCyclodextrin-Mediated Crystallization of Acid Beta-Glucosidase in Complex with Amphiphilic Bicyclic Nojirimycin Analogues.
Brumshtein, B., Aguilar-Moncayo, M., Benito, J.M., Garcia Fernandez, J.M., Silman, I., Shaaltiel, Y., Aviezer, D., Sussman, J.L., Futerman, A.H., Ortiz Mellet, C.(2011) Org Biomol Chem 9: 4160
- PubMed: 21483943
- DOI: https://doi.org/10.1039/c1ob05200d
- Primary Citation of Related Structures:
2XWD, 2XWE - PubMed Abstract:
Cyclodextrin-based host-guest chemistry has been exploited to facilitate co-crystallization of recombinant human acid ¦Â-glucosidase (¦Â-glucocerebrosidase, GlcCerase) with amphiphilic bicyclic nojirimycin analogues of the sp(2)-iminosugar type. Attempts to co-crystallize GlcCerase with 5-N,6-O-[N'-(n-octyl)iminomethylidene]nojirimycin (NOI-NJ) or with 5-N,6-S-[N'-(n-octyl)iminomethylidene]-6-thionojirimycin (6S-NOI-NJ), two potent inhibitors of the enzyme with promising pharmacological chaperone activity for several Gaucher disease-associated mutations, were unsuccessful probably due to the formation of aggregates that increase the heterogeneity of the sample and affect nucleation and growth of crystals. Cyclomaltoheptaose (¦Â-cyclodextrin, ¦ÂCD) efficiently captures NOI-NJ and 6S-NOI-NJ in aqueous media to form inclusion complexes in which the lipophilic tail is accommodated in the hydrophobic cavity of the cyclooligosaccharide. The dissociation constant of the complex of the amphiphilic sp(2)-iminosugars with ¦ÂCD is two orders of magnitude higher than that of the corresponding complex with GlcCerase, allowing the efficient transfer of the inhibitor from the ¦ÂCD cavity to the GlcCerase active site. Enzyme-inhibitor complexes suitable for X-ray analysis were thus grown in the presence of ¦ÂCD. In contrast to what was previously observed for the complex of GlcCerase with the more basic derivative, 6-amino-6-deoxy-5-N,6-N-[N'-(n-octyl)iminomethylidene]nojirimycin (6N-NOI-NJ), the ¦Â-anomers of both NOI-NJ and 6S-NOI-NJ were seen in the active site, even though the ¦Á-anomer was exclusively detected both in aqueous solution and in the corresponding ¦ÂCD:sp(2)-iminosugar complexes. Our results further suggest that cyclodextrin derivatives might serve as suitable delivery systems of amphiphilic glycosidase inhibitors in a biomedical context.
Organizational Affiliation:
Department of Structural Biology, Weizmann Institute of Science, Rehovot, 76100, Israel.
