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2020-04-07Author
Manschwetus, Jascha TobiasWallbott, MaximilianFachinger, AlexandraObergruber, ClaudiaPautz, SabineBertinetti, DanielaSchmidt, Sven H.Herberg, Friedrich W.Subject
540 Chemistry 570 Life sciences; biology 610 Medicine and health Parkinson-KrankheitProtein-Serin-Threonin-KinasenPhosphorylierungMetadata
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Aufsatz
Binding of the Human 14-3-3 Isoforms to Distinct Sites in the Leucine-Rich Repeat Kinase 2
Abstract
Proteins of the 14-3-3 family are well known modulators of the leucine-rich repeat kinase 2 (LRRK2) regulating kinase activity, cellular localization, and ubiquitylation. Although binding between those proteins has been investigated, a comparative study of all human 14-3-3 isoforms interacting with LRRK2 is lacking so far. In a comprehensive approach, we quantitatively analyzed the interaction between the seven human 14-3-3 isoforms and LRRK2-derived peptides covering both, reported and putative 14-3-3 binding sites. We observed that phosphorylation is an absolute prerequisite for 14-3-3 binding and generated binding patterns of 14-3-3 isoforms to interact with peptides derived from the N-terminal phosphorylation cluster (S910 and S935), the Roc domain (S1444) and the C-terminus. The tested 14-3-3 binding sites in LRRK2 preferentially were recognized by the isoforms γ and η, whereas the isoforms ϵ and especially σ showed the weakest or no binding. Interestingly, the possible pathogenic mutation Q930R in LRRK2 drastically increases binding affinity to a peptide encompassing pS935. We then identified the autophosphorylation site T2524 as a so far not described 14-3-3 binding site at the very C-terminus of LRRK2. Binding affinities of all seven 14-3-3 isoforms were quantified for all three binding regions with pS1444 displaying the highest affinity of all measured singly phosphorylated peptides. The strongest binding was detected for the combined phosphosites S910 and S935, suggesting that avidity effects are important for high affinity interaction between 14-3-3 proteins and LRRK2.
Citation
In: Frontiers in Neuroscience 2020 / Volume 14 (2020-04-07) , S. Article 302 ; ISSN 1662-453XSponsorship
Gefördert durch den Publikationsfonds der Universität KasselCitation
@article{doi:10.17170/kobra-202005291309,
author={Manschwetus, Jascha Tobias and Wallbott, Maximilian and Fachinger, Alexandra and Obergruber, Claudia and Pautz, Sabine and Bertinetti, Daniela and Schmidt, Sven H. and Herberg, Friedrich W.},
title={Binding of the Human 14-3-3 Isoforms to Distinct Sites in the Leucine-Rich Repeat Kinase 2},
journal={Frontiers in Neuroscience},
year={2020}
}
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2020-05-29T07:00:56Z 2020-05-29T07:00:56Z 2020-04-07 doi:10.17170/kobra-202005291309 http://hdl.handle.net/123456789/11576 Gefördert durch den Publikationsfonds der Universität Kassel eng Namensnennung 4.0 International http://creativecommons.org/licenses/by/4.0/ LRRK2 14-3-3 proteins isoform specificity Parkinson’s disease phosphorylation 540 570 610 Binding of the Human 14-3-3 Isoforms to Distinct Sites in the Leucine-Rich Repeat Kinase 2 Aufsatz Proteins of the 14-3-3 family are well known modulators of the leucine-rich repeat kinase 2 (LRRK2) regulating kinase activity, cellular localization, and ubiquitylation. Although binding between those proteins has been investigated, a comparative study of all human 14-3-3 isoforms interacting with LRRK2 is lacking so far. In a comprehensive approach, we quantitatively analyzed the interaction between the seven human 14-3-3 isoforms and LRRK2-derived peptides covering both, reported and putative 14-3-3 binding sites. We observed that phosphorylation is an absolute prerequisite for 14-3-3 binding and generated binding patterns of 14-3-3 isoforms to interact with peptides derived from the N-terminal phosphorylation cluster (S910 and S935), the Roc domain (S1444) and the C-terminus. The tested 14-3-3 binding sites in LRRK2 preferentially were recognized by the isoforms γ and η, whereas the isoforms ϵ and especially σ showed the weakest or no binding. Interestingly, the possible pathogenic mutation Q930R in LRRK2 drastically increases binding affinity to a peptide encompassing pS935. We then identified the autophosphorylation site T2524 as a so far not described 14-3-3 binding site at the very C-terminus of LRRK2. Binding affinities of all seven 14-3-3 isoforms were quantified for all three binding regions with pS1444 displaying the highest affinity of all measured singly phosphorylated peptides. The strongest binding was detected for the combined phosphosites S910 and S935, suggesting that avidity effects are important for high affinity interaction between 14-3-3 proteins and LRRK2. open access Manschwetus, Jascha Tobias Wallbott, Maximilian Fachinger, Alexandra Obergruber, Claudia Pautz, Sabine Bertinetti, Daniela Schmidt, Sven H. Herberg, Friedrich W. doi:10.3389/fnins.2020.00302 Parkinson-Krankheit Protein-Serin-Threonin-Kinasen Phosphorylierung publishedVersion ISSN 1662-453X Volume 14 Frontiers in Neuroscience Article 302 2020 false
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