%0 Chart or Table
%A Gloeckner, Christian Johannes
%T Dataset: Supplemental data for 'Intramolecular feedback regulation of the LRRK2 Roc G domain by a LRRK2 kinase dependent mechanism' (Gilsbach et al., eLife 2024, doi:10.7554/eLife.91083), v2
%I Zenodo
%M DZNE-2024-01346
%D 2024
%X Supportive data for the eLife version of record. (1) Data used for the Michaelis Menten Kinetics. HPLC-based assay. Steady-state kinetic measurements of LRRK2-mediated GTP hydrolysis were performed as previously described (Ahmadian et al., 1997). Briefly, 0.1 µM of full-length LRRK2 was incubated with different amounts of GTP (0, 25, 75, 150, 250, 500, 1000, 2000, 3000 and 5000 µM) and production of GDP was monitored by reversed phase C18 HPLC. To this end, the samples (10 µl) were directly injected on a reversed-phase C18 column (pre-column: Hypersil Gold, 3µm particle size, 4.6x10mm; main column: Hypersil Gold, 5µm particle size, 4.6x250mm, Thermo Scientific) using an Ultimate 3000 HPLC system (Thermo Scientific, Waltham, MA, USA) in HPLC-buffer containing 50 mM KH2PO4/K2HPO4 pH 6.0, 10 mM tetrabutylammonium bromide and 10-15
%K structural modeling (Other)
%K LRRK2 (Other)
%F PUB:(DE-HGF)32
%9 Dataset
%R 10.5281/zenodo.13837193
%U https://pub.dzne.de/record/272967