Biochemical and kinetic properties of the complex Roco G-protein cycleWauters, L., Terheyden, S., Gilsbach, B. K., Leemans, M., Athanasopoulos, P. S., Guaitoli, G., Wittinghofer, A., Gloeckner, C. J., Versées, W. & Kortholt, A., Nov-2018, In : Biological Chemistry. 399, 12, p. 1447-1456 10 p.
Research output: Contribution to journal › Article › Academic › peer-review
Roco proteins have come into focus after mutations in the gene coding for the human Roco protein Leucine-rich repeat kinase 2 (LRRK2) were discovered to be one of the most common genetic causes of late onset Parkinson's disease. Roco proteins are characterized by a Roc domain responsible for GTP binding and hydrolysis, followed by a COR dimerization device. The regulation and function of this RocCOR domain tandem is still not completely understood. To fully biochemically characterize Roco proteins, we performed a systematic survey of the kinetic properties of several Roco protein family members, including LRRK2. Together, our results show that Roco proteins have a unique G-protein cycle. Our results confirm that Roco proteins have a low nucleotide affinity in the micromolar range and thus do not strictly depend on G-nucleotide exchange factors. Measurement of multiple and single turnover reactions shows that neither Pi nor GDP release are rate-limiting, while this is the case for the GAP-mediated GTPase reaction of some small G-proteins like Ras and for most other high affinity Ras-like proteins, respectively. The KM values of the reactions are in the range of the physiological GTP concentration, suggesting that LRRK2 functioning might be regulated by the cellular GTP level.
|Number of pages||10|
|Early online date||28-Jul-2018|
|Publication status||Published - Nov-2018|
- GTPase, leucine-rich repeat kinase 2, Roco proteins, unconventional G-protein