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Conformational heterogeneity of the Roc domains in C. tepidum Roc-COR and implications for human LRRK2 Parkinson mutations

Rudi, K., Ho, F. Y., Gilsbach, B. K., Pots, H., Wittinghofer, A., Kortholt, A. & Klare, J. P., Oct-2015, In : Bioscience reports. 35, 5, p. 1-12 12 p., e00254.

Research output: Contribution to journalArticleAcademicpeer-review

Ras of complex proteins (Roc) is a Ras-like GTP binding domain that always occurs in tandem with the C-terminal of Roc (COR) domain, and is found in bacteria, plants and animals. Recently, it has been shown that Roco proteins belong to the family of G-proteins activated by nucleotide-dependent dimerization (GADs). We investigated the RocCOR tandem from the bacteria Chlorobium tepidum with site-directed spin labeling and pulse EPR distance measurements to follow conformational changes during the Roco G-protein cycle. Our results confirm that the COR domains are a stable dimerization device serving as a scaffold for the Roc domains, that in contrast are structurally heterogeneous and dynamic entities. Contrary to other GAD proteins, we observed only minor structural alterations upon binding and hydrolysis of GTP, indicating significant mechanistic variations within this protein class. Mutations in the most prominent member of the Roco family of proteins, leucine-rich repeat kinase 2 (LRRK2), are the most frequent cause of late-onset Parkinson's disease (PD). Using a stable recombinant LRRK2 Roc-COR-Kinase fragment we obtained detailed kinetic data for the G-protein cycle. Our data confirmed that dimerization is essential for efficient GTP hydrolysis, and PD mutations in the Roc domain result in decreased GTPase activity. Previous data have shown that these LRRK2 PD-mutations are located in the interface between Roc and COR. Importantly, analogous mutations in the conserved C. tepidum RocCOR interface significantly influence the structure and nucleotide-induced conformational changes of the Roc domains.

Original languageEnglish
Article numbere00254
Pages (from-to)1-12
Number of pages12
JournalBioscience reports
Volume35
Issue number5
Publication statusPublished - Oct-2015

    Keywords

  • ELECTRON-ELECTRON RESONANCE, LEUCINE-RICH-REPEAT-KINASE-2 LRRK2, COMPLEX PROTEINS, GTPASE ACTIVITY, KINASE, DIMERIZATION, INSIGHTS, CYTOSKELETON, MUTANTS

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