Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.

Human dynamin-1-like protein (DNM1L) is a GTP-driven molecular machine that segregates mitochondria and peroxisomes. To obtain insights into its catalytic mechanism, we determined crystal structures of a construct comprising the GTPase domain and the bundle signaling element (BSE) in the nucleotide-...

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Autores principales:	Julia Wenger, Eva Klinglmayr, Chris Fröhlich, Clarissa Eibl, Ana Gimeno, Manuel Hessenberger, Sandra Puehringer, Oliver Daumke, Peter Goettig
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6e3cec825e404f56a4b6e73fabce42dc
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spelling	oai:doaj.org-article:6e3cec825e404f56a4b6e73fabce42dc2021-11-18T08:59:06ZFunctional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.1932-620310.1371/journal.pone.0071835https://doaj.org/article/6e3cec825e404f56a4b6e73fabce42dc2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23977156/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Human dynamin-1-like protein (DNM1L) is a GTP-driven molecular machine that segregates mitochondria and peroxisomes. To obtain insights into its catalytic mechanism, we determined crystal structures of a construct comprising the GTPase domain and the bundle signaling element (BSE) in the nucleotide-free and GTP-analogue-bound states. The GTPase domain of DNM1L is structurally related to that of dynamin and binds the nucleotide 5'-Guanylyl-imidodiphosphate (GMP-PNP) via five highly conserved motifs, whereas the BSE folds into a pocket at the opposite side. Based on these structures, the GTPase center was systematically mapped by alanine mutagenesis and kinetic measurements. Thus, residues essential for the GTPase reaction were characterized, among them Lys38, Ser39 and Ser40 in the phosphate binding loop, Thr59 from switch I, Asp146 and Gly149 from switch II, Lys216 and Asp218 in the G4 element, as well as Asn246 in the G5 element. Also, mutated Glu81 and Glu82 in the unique 16-residue insertion of DNM1L influence the activity significantly. Mutations of Gln34, Ser35, and Asp190 in the predicted assembly interface interfered with dimerization of the GTPase domain induced by a transition state analogue and led to a loss of the lipid-stimulated GTPase activity. Our data point to related catalytic mechanisms of DNM1L and dynamin involving dimerization of their GTPase domains.Julia WengerEva KlinglmayrChris FröhlichClarissa EiblAna GimenoManuel HessenbergerSandra PuehringerOliver DaumkePeter GoettigPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e71835 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Julia Wenger Eva Klinglmayr Chris Fröhlich Clarissa Eibl Ana Gimeno Manuel Hessenberger Sandra Puehringer Oliver Daumke Peter Goettig Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.
description	Human dynamin-1-like protein (DNM1L) is a GTP-driven molecular machine that segregates mitochondria and peroxisomes. To obtain insights into its catalytic mechanism, we determined crystal structures of a construct comprising the GTPase domain and the bundle signaling element (BSE) in the nucleotide-free and GTP-analogue-bound states. The GTPase domain of DNM1L is structurally related to that of dynamin and binds the nucleotide 5'-Guanylyl-imidodiphosphate (GMP-PNP) via five highly conserved motifs, whereas the BSE folds into a pocket at the opposite side. Based on these structures, the GTPase center was systematically mapped by alanine mutagenesis and kinetic measurements. Thus, residues essential for the GTPase reaction were characterized, among them Lys38, Ser39 and Ser40 in the phosphate binding loop, Thr59 from switch I, Asp146 and Gly149 from switch II, Lys216 and Asp218 in the G4 element, as well as Asn246 in the G5 element. Also, mutated Glu81 and Glu82 in the unique 16-residue insertion of DNM1L influence the activity significantly. Mutations of Gln34, Ser35, and Asp190 in the predicted assembly interface interfered with dimerization of the GTPase domain induced by a transition state analogue and led to a loss of the lipid-stimulated GTPase activity. Our data point to related catalytic mechanisms of DNM1L and dynamin involving dimerization of their GTPase domains.
format	article
author	Julia Wenger Eva Klinglmayr Chris Fröhlich Clarissa Eibl Ana Gimeno Manuel Hessenberger Sandra Puehringer Oliver Daumke Peter Goettig
author_facet	Julia Wenger Eva Klinglmayr Chris Fröhlich Clarissa Eibl Ana Gimeno Manuel Hessenberger Sandra Puehringer Oliver Daumke Peter Goettig
author_sort	Julia Wenger
title	Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.
title_short	Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.
title_full	Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.
title_fullStr	Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.
title_full_unstemmed	Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.
title_sort	functional mapping of human dynamin-1-like gtpase domain based on x-ray structure analyses.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/6e3cec825e404f56a4b6e73fabce42dc
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_version_	1718421088129515520

Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses.

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