Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes.
Details
Serval ID
serval:BIB_1529E84C68A5
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes.
Journal
Scientific reports
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Publication state
Published
Issued date
17/08/2017
Peer-reviewed
Oui
Volume
7
Number
1
Pages
8667
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Publication Status: epublish
Abstract
Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain protein implicated in the pathogenesis of both familial and sporadic Parkinson's disease (PD), and currently one of the most promising therapeutic targets for drug design in Parkinson's disease. In contrast, LRRK1, the closest homologue to LRRK2, does not play any role in PD. Here, we use cryo-electron microscopy (cryo-EM) and single particle analysis to gain structural insight into the full-length dimeric structures of LRRK2 and LRRK1. Differential scanning fluorimetry-based screening of purification buffers showed that elution of the purified LRRK2 protein in a high pH buffer is beneficial in obtaining high quality cryo-EM images. Next, analysis of the 3D maps generated from the cryo-EM data show 16 and 25 Å resolution structures of full length LRRK2 and LRRK1, respectively, revealing the overall shape of the dimers with two-fold symmetric orientations of the protomers that is closely similar between the two proteins. These results suggest that dimerization mechanisms of both LRRKs are closely related and hence that specificities in functions of each LRRK are likely derived from LRRK2 and LRRK1's other biochemical functions. To our knowledge, this study is the first to provide 3D structural insights in LRRK2 and LRRK1 dimers in parallel.
Keywords
Buffers, Cryoelectron Microscopy, Detergents/pharmacology, Humans, Imaging, Three-Dimensional, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/chemistry, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/metabolism, Protein Folding/drug effects, Protein Multimerization/drug effects, Protein Serine-Threonine Kinases/chemistry, Protein Serine-Threonine Kinases/metabolism, Solutions
Pubmed
Web of science
Open Access
Yes
Create date
09/06/2023 15:02
Last modification date
08/07/2023 5:50