Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties
Cathepsin D (CTSD) is a lysosomal protease important for the degradation of various substrates, including disease-associated proteins like α-synuclein (a-syn), amyloid precursor protein (APP) and tau, all of which tend to aggregate if not efficiently degraded. Hence, it is not surprising that geneti...
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| Format: | Article |
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Frontiers Media S.A.
2021-02-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2021.581805/full |
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| author | Josina Bunk Susy Prieto Huarcaya Susy Prieto Huarcaya Alice Drobny Alice Drobny Jan Philipp Dobert Jan Philipp Dobert Lina Walther Stefan Rose-John Philipp Arnold Philipp Arnold Friederike Zunke Friederike Zunke |
| author_facet | Josina Bunk Susy Prieto Huarcaya Susy Prieto Huarcaya Alice Drobny Alice Drobny Jan Philipp Dobert Jan Philipp Dobert Lina Walther Stefan Rose-John Philipp Arnold Philipp Arnold Friederike Zunke Friederike Zunke |
| author_sort | Josina Bunk |
| collection | DOAJ |
| description | Cathepsin D (CTSD) is a lysosomal protease important for the degradation of various substrates, including disease-associated proteins like α-synuclein (a-syn), amyloid precursor protein (APP) and tau, all of which tend to aggregate if not efficiently degraded. Hence, it is not surprising that genetic variants within the CTSD gene have been linked to neurodegenerative diseases, like Parkinson’s and Alzheimer’s disease (PD, AD), as well as the lysosomal storage disorder neuronal ceroid lipofuscinosis type-10 (NCL10). Although recent studies have shown the molecular dependence of substrate degradation via CTSD within autophagic pathways, only little is known about the precise role of lysosomal CTSD function in disease development. We here performed biochemical, cellular and structural analyses of eleven disease-causing CTSD point mutations found in genomic sequencing data of patients to understand their role in neurodegeneration. These CTSD variants were analyzed for cellular localization, maturation and enzymatic activity in overexpression analyses. Moreover, for PD-associated mutants, intracellular degradation of a-syn was monitored. In summary, our results suggest that NCL10-associated CTSD variants are significantly impaired in lysosomal maturation and enzymatic activity, whereas the AD- and PD-associated variants seemed rather unaffected, indicating normal maturation, and lysosomal presence. Interestingly, a PD-associated CTSD variant (A239V) exhibited increased enzymatic activity accompanied by enhanced a-syn degradation. By structural analyses of this mutant utilizing molecular dynamics simulation (MDS), we identified a structural change within a loop adjacent to the catalytic center leading to a higher flexibility and potentially accelerated substrate exchange rates. Our data sheds light onto the role of CTSD in disease development and helps to understand the structural regulation of enzymatic function, which could be utilized for targeted CTSD activation. Because of the degradative function of CTSD, this enzyme is especially interesting for therapeutic strategies tackling protein aggregates in neurodegenerative disorders. |
| first_indexed | 2024-12-19T17:19:04Z |
| format | Article |
| id | doaj.art-6862f2b9aeb04f2b90dddd37967cda70 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-12-19T17:19:04Z |
| publishDate | 2021-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-6862f2b9aeb04f2b90dddd37967cda702022-12-21T20:12:45ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2021-02-01910.3389/fcell.2021.581805581805Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation PropertiesJosina Bunk0Susy Prieto Huarcaya1Susy Prieto Huarcaya2Alice Drobny3Alice Drobny4Jan Philipp Dobert5Jan Philipp Dobert6Lina Walther7Stefan Rose-John8Philipp Arnold9Philipp Arnold10Friederike Zunke11Friederike Zunke12Institute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyInstitute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyDepartment of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyInstitute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyDepartment of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyInstitute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyDepartment of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyInstitute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyInstitute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyInstitute of Anatomy, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyInstitute of Anatomy, Functional and Clinical Anatomy, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, GermanyInstitute of Biochemistry, Christian-Albrechts-Universität zu Kiel, Kiel, GermanyDepartment of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyCathepsin D (CTSD) is a lysosomal protease important for the degradation of various substrates, including disease-associated proteins like α-synuclein (a-syn), amyloid precursor protein (APP) and tau, all of which tend to aggregate if not efficiently degraded. Hence, it is not surprising that genetic variants within the CTSD gene have been linked to neurodegenerative diseases, like Parkinson’s and Alzheimer’s disease (PD, AD), as well as the lysosomal storage disorder neuronal ceroid lipofuscinosis type-10 (NCL10). Although recent studies have shown the molecular dependence of substrate degradation via CTSD within autophagic pathways, only little is known about the precise role of lysosomal CTSD function in disease development. We here performed biochemical, cellular and structural analyses of eleven disease-causing CTSD point mutations found in genomic sequencing data of patients to understand their role in neurodegeneration. These CTSD variants were analyzed for cellular localization, maturation and enzymatic activity in overexpression analyses. Moreover, for PD-associated mutants, intracellular degradation of a-syn was monitored. In summary, our results suggest that NCL10-associated CTSD variants are significantly impaired in lysosomal maturation and enzymatic activity, whereas the AD- and PD-associated variants seemed rather unaffected, indicating normal maturation, and lysosomal presence. Interestingly, a PD-associated CTSD variant (A239V) exhibited increased enzymatic activity accompanied by enhanced a-syn degradation. By structural analyses of this mutant utilizing molecular dynamics simulation (MDS), we identified a structural change within a loop adjacent to the catalytic center leading to a higher flexibility and potentially accelerated substrate exchange rates. Our data sheds light onto the role of CTSD in disease development and helps to understand the structural regulation of enzymatic function, which could be utilized for targeted CTSD activation. Because of the degradative function of CTSD, this enzyme is especially interesting for therapeutic strategies tackling protein aggregates in neurodegenerative disorders.https://www.frontiersin.org/articles/10.3389/fcell.2021.581805/fulllysosomal degradationmolecular dynamics simulationParkinson’s diseaseneuronal ceroid lipofuscinoseslysosomesalpha-synuclein |
| spellingShingle | Josina Bunk Susy Prieto Huarcaya Susy Prieto Huarcaya Alice Drobny Alice Drobny Jan Philipp Dobert Jan Philipp Dobert Lina Walther Stefan Rose-John Philipp Arnold Philipp Arnold Friederike Zunke Friederike Zunke Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties Frontiers in Cell and Developmental Biology lysosomal degradation molecular dynamics simulation Parkinson’s disease neuronal ceroid lipofuscinoses lysosomes alpha-synuclein |
| title | Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties |
| title_full | Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties |
| title_fullStr | Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties |
| title_full_unstemmed | Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties |
| title_short | Cathepsin D Variants Associated With Neurodegenerative Diseases Show Dysregulated Functionality and Modified α-Synuclein Degradation Properties |
| title_sort | cathepsin d variants associated with neurodegenerative diseases show dysregulated functionality and modified α synuclein degradation properties |
| topic | lysosomal degradation molecular dynamics simulation Parkinson’s disease neuronal ceroid lipofuscinoses lysosomes alpha-synuclein |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2021.581805/full |
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