The Roc-COR tandem domain of leucine-rich repeat kinase 2 forms dimers and exhibits conventional Ras-like GTPase properties
| Autor: | George Cao, Daisy Sio-Seng Lio, Michael D. W. Griffin, Ryan D. Mills, Lung-Yu Liang, Janetta G. Culvenor, Heung-Chin Cheng, Vijaya Kenche, Terrence D. Mulhern, Yee-Foong Mok |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
rac1 GTP-Binding Protein
0301 basic medicine GTP' Guanine Stereochemistry G protein GTPase Leucine-rich repeat Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 Biochemistry Gene Expression Regulation Enzymologic Protein Structure Secondary GTP Phosphohydrolases Mice 03 medical and health sciences Cellular and Molecular Neuroscience chemistry.chemical_compound 0302 clinical medicine Protein structure Escherichia coli Animals Humans Magnesium Kinase Neuropeptides LRRK2 Guanine Nucleotides Recombinant Proteins nervous system diseases Genes ras 030104 developmental biology chemistry Mutation Protein Multimerization Dimerization 030217 neurology & neurosurgery |
| Zdroj: | Journal of Neurochemistry. 147:409-428 |
| ISSN: | 0022-3042 |
| Popis: | The Parkinson's disease (PD)-causative leucine-rich repeat kinase 2 (LRRK2) belongs to the Roco family of G-proteins comprising a Ras-of-complex (Roc) domain followed by a C-terminal of Roc (COR) domain in tandem (called Roc-COR domain). Two prokaryotic Roc-COR domains have been characterized as 'G proteins activated by guanine nucleotide-dependent dimerization' (GADs), which require dimerization for activation of their GTPase activity and bind guanine nucleotides with relatively low affinities. Additionally, LRRK2 Roc domain in isolation binds guanine nucleotides with relatively low affinities. As such, LRRK2 GTPase domain was predicted to be a GAD. Herein, we describe the design and high-level expression of human LRRK2 Roc-COR domain (LRRK2 Roc-COR). Biochemical analyses of LRRK2 Roc-COR reveal that it forms homodimers, with the C-terminal portion of COR mediating its dimerization. Furthermore, it co-purifies and binds Mg2+ GTP/GDP at 1 : 1 stoichiometry, and it hydrolyzes GTP with Km and kcat of 22 nM and 4.70 × 10-4 min-1 , respectively. Thus, even though LRRK2 Roc-COR forms GAD-like homodimers, it exhibits conventional Ras-like GTPase properties, with high-affinity binding of Mg2+ -GTP/GDP and low intrinsic catalytic activity. The PD-causative Y1699C mutation mapped to the COR domain was previously reported to reduce the GTPase activity of full-length LRRK2. In contrast, this mutation induces no change in the GTPase activity, and only slight perturbations in the secondary structure contents of LRRK2 Roc-COR. As this mutation does not directly affect the GTPase activity of the isolated Roc-COR tandem, it is possible that the effects of this mutation on full-length LRRK2 occur via other functional domains. Open Practices Open Science: This manuscript was awarded with the Open Materials Badge. For more information see: https://cos.io/our-services/open-science-badges/. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text