Article
Cyclodextrin-mediated Crystallization of Acid β-glucosidase in Complex with Amphiphilic Bicyclic Nojirimycin Analogues
Author/s | Brumshtein, Boris
Aguilar Moncayo, Matilde Benito, Juan Manuel García Fernández, José Manuel Silman, Israel Shaaltiel, Yoseph Aviezer, David Sussman, Joel L. Futerman, Anthony H. Ortiz Mellet, Carmen |
Department | Universidad de Sevilla. Departamento de Química orgánica |
Publication Date | 2011 |
Deposit Date | 2022-11-11 |
Published in |
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Abstract | Cyclodextrin-based host-guest chemistry has been exploited to facilitate co-crystallization of recombinant human acid β-glucosidase (β-glucocerebrosidase, GlcCerase) with amphiphilic bicyclic nojirimycin analogues of the ... Cyclodextrin-based host-guest chemistry has been exploited to facilitate co-crystallization of recombinant human acid β-glucosidase (β-glucocerebrosidase, GlcCerase) with amphiphilic bicyclic nojirimycin analogues of the sp2-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 (β-cyclodextrin, βCD) 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 sp2-iminosugars with βCD is two orders of magnitude higher than that of the corresponding complex with GlcCerase, allowing the efficient transfer of the inhibitor from the βCD cavity to the GlcCerase active site. Enzyme-inhibitor complexes suitable for X-ray analysis were thus grown in the presence of βCD. 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 β-anomers of both NOI-NJ and 6S-NOI-NJ were seen in the active site, even though the α-anomer was exclusively detected both in aqueous solution and in the corresponding βCD:sp2-iminosugar complexes. Our results further suggest that cyclodextrin derivatives might serve as suitable delivery systems of amphiphilic glycosidase inhibitors in a biomedical context. |
Funding agencies | Ministerio de Ciencia e Innovación (MICIN). España Junta de Andalucía Fondo Europeo de Desarrollo Regional |
Project ID. | CTQ2007-61180/PPQ
SAF2010-15670 CTQ2010-15848 P08-FQM-03711 03122 ISSG-CT-2007-037198 |
Citation | Brumshtein, B., Aguilar Moncayo, M., Benito, J.M., García Fernández, J.M., Silman, I., Shaaltiel, Y.,...,Ortiz Mellet, C. (2011). Cyclodextrin-mediated Crystallization of Acid β-glucosidase in Complex with Amphiphilic Bicyclic Nojirimycin Analogues. Organic and Biomolecular Chemistry, 9 (11), 4160-4167. https://doi.org/10.1039/c1ob05200d. |
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