Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function
Abstract The enzyme glucocerebrosidase (GCase) catalyses the hydrolysis of glucosylceramide to glucose and ceramide within lysosomes. Homozygous or compound heterozygous mutations in the GCase-encoding GBA1 gene cause the lysosomal storage disorder Gaucher disease, while heterozygous and homozygous...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-05-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-60134-4 |
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| author | Thomas Dal Maso Chiara Sinisgalli Gianluca Zilio Elisa Franzin Isabella Tessari Els Pardon Jan Steyaert Steven Ballet Elisa Greggio Wim Versées Nicoletta Plotegher |
| author_facet | Thomas Dal Maso Chiara Sinisgalli Gianluca Zilio Elisa Franzin Isabella Tessari Els Pardon Jan Steyaert Steven Ballet Elisa Greggio Wim Versées Nicoletta Plotegher |
| author_sort | Thomas Dal Maso |
| collection | DOAJ |
| description | Abstract The enzyme glucocerebrosidase (GCase) catalyses the hydrolysis of glucosylceramide to glucose and ceramide within lysosomes. Homozygous or compound heterozygous mutations in the GCase-encoding GBA1 gene cause the lysosomal storage disorder Gaucher disease, while heterozygous and homozygous mutations are the most frequent genetic risk factor for Parkinson’s disease. These mutations commonly affect GCase stability, trafficking or activity. Here, we report the development and characterization of nanobodies (Nbs) targeting and acting as molecular chaperones for GCase. We identify several Nb families that bind with nanomolar affinity to GCase. Based on biochemical characterization, we group the Nbs in two classes: Nbs that improve the activity of the enzyme and Nbs that increase GCase stability in vitro. A selection of the most promising Nbs is shown to improve GCase function in cell models and positively impact the activity of the N370S mutant GCase. These results lay the foundation for the development of new therapeutic routes. |
| format | Article |
| id | doaj-art-bfb317ed106347bebcca7900f74c935c |
| institution | OA Journals |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-bfb317ed106347bebcca7900f74c935c2025-08-20T02:03:31ZengNature PortfolioNature Communications2041-17232025-05-0116111910.1038/s41467-025-60134-4Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase functionThomas Dal Maso0Chiara Sinisgalli1Gianluca Zilio2Elisa Franzin3Isabella Tessari4Els Pardon5Jan Steyaert6Steven Ballet7Elisa Greggio8Wim Versées9Nicoletta Plotegher10VIB-VUB Center for Structural Biology, VIBDepartment of Biology, University of PadovaDepartment of Biology, University of PadovaDepartment of Biology, University of PadovaDepartment of Biology, University of PadovaVIB-VUB Center for Structural Biology, VIBVIB-VUB Center for Structural Biology, VIBResearch Group of Organic Chemistry, Vrije Universiteit BrusselDepartment of Biology, University of PadovaVIB-VUB Center for Structural Biology, VIBDepartment of Biology, University of PadovaAbstract The enzyme glucocerebrosidase (GCase) catalyses the hydrolysis of glucosylceramide to glucose and ceramide within lysosomes. Homozygous or compound heterozygous mutations in the GCase-encoding GBA1 gene cause the lysosomal storage disorder Gaucher disease, while heterozygous and homozygous mutations are the most frequent genetic risk factor for Parkinson’s disease. These mutations commonly affect GCase stability, trafficking or activity. Here, we report the development and characterization of nanobodies (Nbs) targeting and acting as molecular chaperones for GCase. We identify several Nb families that bind with nanomolar affinity to GCase. Based on biochemical characterization, we group the Nbs in two classes: Nbs that improve the activity of the enzyme and Nbs that increase GCase stability in vitro. A selection of the most promising Nbs is shown to improve GCase function in cell models and positively impact the activity of the N370S mutant GCase. These results lay the foundation for the development of new therapeutic routes.https://doi.org/10.1038/s41467-025-60134-4 |
| spellingShingle | Thomas Dal Maso Chiara Sinisgalli Gianluca Zilio Elisa Franzin Isabella Tessari Els Pardon Jan Steyaert Steven Ballet Elisa Greggio Wim Versées Nicoletta Plotegher Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function Nature Communications |
| title | Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function |
| title_full | Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function |
| title_fullStr | Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function |
| title_full_unstemmed | Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function |
| title_short | Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function |
| title_sort | developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function |
| url | https://doi.org/10.1038/s41467-025-60134-4 |
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