Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes

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 homo...

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Autores principales:	Kushal Sejwal, Mohamed Chami, Hervé Rémigy, Renée Vancraenenbroeck, William Sibran, Rosmarie Sütterlin, Paul Baumgartner, Robert McLeod, Marie-Christine Chartier-Harlin, Veerle Baekelandt, Henning Stahlberg, Jean-Marc Taymans
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b9e5473e709a4a3ca47905077e0c271d
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spelling	oai:doaj.org-article:b9e5473e709a4a3ca47905077e0c271d2021-12-02T12:31:45ZCryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes10.1038/s41598-017-09126-z2045-2322https://doaj.org/article/b9e5473e709a4a3ca47905077e0c271d2017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-09126-zhttps://doaj.org/toc/2045-2322Abstract 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.Kushal SejwalMohamed ChamiHervé RémigyRenée VancraenenbroeckWilliam SibranRosmarie SütterlinPaul BaumgartnerRobert McLeodMarie-Christine Chartier-HarlinVeerle BaekelandtHenning StahlbergJean-Marc TaymansNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kushal Sejwal Mohamed Chami Hervé Rémigy Renée Vancraenenbroeck William Sibran Rosmarie Sütterlin Paul Baumgartner Robert McLeod Marie-Christine Chartier-Harlin Veerle Baekelandt Henning Stahlberg Jean-Marc Taymans Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes
description	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.
format	article
author	Kushal Sejwal Mohamed Chami Hervé Rémigy Renée Vancraenenbroeck William Sibran Rosmarie Sütterlin Paul Baumgartner Robert McLeod Marie-Christine Chartier-Harlin Veerle Baekelandt Henning Stahlberg Jean-Marc Taymans
author_facet	Kushal Sejwal Mohamed Chami Hervé Rémigy Renée Vancraenenbroeck William Sibran Rosmarie Sütterlin Paul Baumgartner Robert McLeod Marie-Christine Chartier-Harlin Veerle Baekelandt Henning Stahlberg Jean-Marc Taymans
author_sort	Kushal Sejwal
title	Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes
title_short	Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes
title_full	Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes
title_fullStr	Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes
title_full_unstemmed	Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes
title_sort	cryo-em analysis of homodimeric full-length lrrk2 and lrrk1 protein complexes
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/b9e5473e709a4a3ca47905077e0c271d
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Cryo-EM analysis of homodimeric full-length LRRK2 and LRRK1 protein complexes

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