Cargando…

Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations

Ras of complex proteins (Roc) is a Ras-like GTP-binding domain that always occurs in tandem with the C-terminal of Roc (COR) domain and is found in bacteria, plants and animals. Recently, it has been shown that Roco proteins belong to the family of G-proteins activated by nucleotide (nt)-dependent d...

Descripción completa

Detalles Bibliográficos
Autores principales: Rudi, Katharina, Ho, Franz Y., Gilsbach, Bernd K., Pots, Henderikus, Wittinghofer, Alfred, Kortholt, Arjan, Klare, Johann P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4721548/
https://www.ncbi.nlm.nih.gov/pubmed/26310572
http://dx.doi.org/10.1042/BSR20150128
_version_ 1782411242196959232
author Rudi, Katharina
Ho, Franz Y.
Gilsbach, Bernd K.
Pots, Henderikus
Wittinghofer, Alfred
Kortholt, Arjan
Klare, Johann P.
author_facet Rudi, Katharina
Ho, Franz Y.
Gilsbach, Bernd K.
Pots, Henderikus
Wittinghofer, Alfred
Kortholt, Arjan
Klare, Johann P.
author_sort Rudi, Katharina
collection PubMed
description Ras of complex proteins (Roc) is a Ras-like GTP-binding domain that always occurs in tandem with the C-terminal of Roc (COR) domain and is found in bacteria, plants and animals. Recently, it has been shown that Roco proteins belong to the family of G-proteins activated by nucleotide (nt)-dependent dimerization (GADs). We investigated the RocCOR tandem from the bacteria Chlorobium tepidum with site-directed spin labelling and pulse EPR distance measurements to follow conformational changes during the Roco G-protein cycle. Our results confirm that the COR domains are a stable dimerization device serving as a scaffold for the Roc domains that, in contrast, are structurally heterogeneous and dynamic entities. Contrary to other GAD proteins, we observed only minor structural alterations upon binding and hydrolysis of GTP, indicating significant mechanistic variations within this protein class. Mutations in the most prominent member of the Roco family of proteins, leucine-rich repeat (LRR) kinase 2 (LRRK2), are the most frequent cause of late-onset Parkinson's disease (PD). Using a stable recombinant LRRK2 Roc-COR-kinase fragment we obtained detailed kinetic data for the G-protein cycle. Our data confirmed that dimerization is essential for efficient GTP hydrolysis and PD mutations in the Roc domain result in decreased GTPase activity. Previous data have shown that these LRRK2 PD-mutations are located in the interface between Roc and COR. Importantly, analogous mutations in the conserved C. tepidum Roc/COR interface significantly influence the structure and nt-induced conformational changes of the Roc domains.
format Online
Article
Text
id pubmed-4721548
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Portland Press Ltd.
record_format MEDLINE/PubMed
spelling pubmed-47215482016-02-02 Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations Rudi, Katharina Ho, Franz Y. Gilsbach, Bernd K. Pots, Henderikus Wittinghofer, Alfred Kortholt, Arjan Klare, Johann P. Biosci Rep Original Papers Ras of complex proteins (Roc) is a Ras-like GTP-binding domain that always occurs in tandem with the C-terminal of Roc (COR) domain and is found in bacteria, plants and animals. Recently, it has been shown that Roco proteins belong to the family of G-proteins activated by nucleotide (nt)-dependent dimerization (GADs). We investigated the RocCOR tandem from the bacteria Chlorobium tepidum with site-directed spin labelling and pulse EPR distance measurements to follow conformational changes during the Roco G-protein cycle. Our results confirm that the COR domains are a stable dimerization device serving as a scaffold for the Roc domains that, in contrast, are structurally heterogeneous and dynamic entities. Contrary to other GAD proteins, we observed only minor structural alterations upon binding and hydrolysis of GTP, indicating significant mechanistic variations within this protein class. Mutations in the most prominent member of the Roco family of proteins, leucine-rich repeat (LRR) kinase 2 (LRRK2), are the most frequent cause of late-onset Parkinson's disease (PD). Using a stable recombinant LRRK2 Roc-COR-kinase fragment we obtained detailed kinetic data for the G-protein cycle. Our data confirmed that dimerization is essential for efficient GTP hydrolysis and PD mutations in the Roc domain result in decreased GTPase activity. Previous data have shown that these LRRK2 PD-mutations are located in the interface between Roc and COR. Importantly, analogous mutations in the conserved C. tepidum Roc/COR interface significantly influence the structure and nt-induced conformational changes of the Roc domains. Portland Press Ltd. 2015-10-08 /pmc/articles/PMC4721548/ /pubmed/26310572 http://dx.doi.org/10.1042/BSR20150128 Text en © 2015 Authors http://creativecommons.org/licenses/by/3.0/ This is an open access article published by Portland Press Limited and distributed under the Creative Commons Attribution Licence 3.0 (http://creativecommons.org/licenses/by/3.0/) .
spellingShingle Original Papers
Rudi, Katharina
Ho, Franz Y.
Gilsbach, Bernd K.
Pots, Henderikus
Wittinghofer, Alfred
Kortholt, Arjan
Klare, Johann P.
Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations
title Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations
title_full Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations
title_fullStr Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations
title_full_unstemmed Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations
title_short Conformational heterogeneity of the Roc domains in C. tepidum Roc–COR and implications for human LRRK2 Parkinson mutations
title_sort conformational heterogeneity of the roc domains in c. tepidum roc–cor and implications for human lrrk2 parkinson mutations
topic Original Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4721548/
https://www.ncbi.nlm.nih.gov/pubmed/26310572
http://dx.doi.org/10.1042/BSR20150128
work_keys_str_mv AT rudikatharina conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations
AT hofranzy conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations
AT gilsbachberndk conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations
AT potshenderikus conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations
AT wittinghoferalfred conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations
AT kortholtarjan conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations
AT klarejohannp conformationalheterogeneityoftherocdomainsinctepidumroccorandimplicationsforhumanlrrk2parkinsonmutations