DC Element | Wert | Sprache |
dc.contributor.author | Meyer, Simon | |
dc.contributor.author | Boehme, Sabine | |
dc.contributor.author | Krueger, Andre | |
dc.contributor.author | Steinhoff, Heinz-Juergen | |
dc.contributor.author | Klare, Johann P. | |
dc.contributor.author | Wittinghofer, Alfred | |
dc.date.accessioned | 2021-12-23T16:11:25Z | - |
dc.date.available | 2021-12-23T16:11:25Z | - |
dc.date.issued | 2009 | |
dc.identifier.issn | 15449173 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/9696 | - |
dc.description.abstract | MnmE, which is involved in the modification of the wobble position of certain tRNAs, belongs to the expanding class of G proteins activated by nucleotide-dependent dimerization (GADs). Previous models suggested the protein to be a multidomain protein whose G domains contact each other in a nucleotide dependent manner. Here we employ a combined approach of X-ray crystallography and pulse electron paramagnetic resonance (EPR) spectroscopy to show that large domain movements are coupled to the G protein cycle of MnmE. The X-ray structures show MnmE to be a constitutive homodimer where the highly mobile G domains face each other in various orientations but are not in close contact as suggested by the GDP-AlF(x) structure of the isolated domains. Distance measurements by pulse double electron-electron resonance (DEER) spectroscopy show that the G domains adopt an open conformation in the nucleotide free/GDP-bound and an open/closed two-state equilibrium in the GTP-bound state, with maximal distance variations of 18 angstrom. With GDP and AlF(x), which mimic the transition state of the phosphoryl transfer reaction, only the closed conformation is observed. Dimerization of the active sites with GDP-AlF(x) requires the presence of specific monovalent cations, thus reflecting the requirements for the GTPase reaction of MnmE. Our results directly demonstrate the nature of the conformational changes MnmE was previously suggested to undergo during its GTPase cycle. They show the nucleotide-dependent dynamic movements of the G domains around two swivel positions relative to the rest of the protein, and they are of crucial importance for understanding the mechanistic principles of this GAD. | |
dc.description.sponsorship | Fond der Chemischen IndustrieFonds der Chemischen Industrie; International Max Planck Research School; Deutsche ForschungsgemeinschaftGerman Research Foundation (DFG) [STE640/7-2]; Graduate College [612]; SM was funded by the Fond der Chemischen Industrie and by the International Max Planck Research School with predoctoral fellowships. SB, JPK, and H-JS are funded by the Deutsche Forschungsgemeinschaft (STE640/7-2) and the Graduate College 612 ( SB). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. | |
dc.language.iso | en | |
dc.publisher | PUBLIC LIBRARY SCIENCE | |
dc.relation.ispartof | PLOS BIOLOGY | |
dc.subject | A1555G MUTATION | |
dc.subject | Biochemistry & Molecular Biology | |
dc.subject | Biology | |
dc.subject | CHAPERONE MACHINERY | |
dc.subject | DIMERIZATION | |
dc.subject | EPR SPECTROSCOPY | |
dc.subject | ESCHERICHIA-COLI | |
dc.subject | EXPRESSION | |
dc.subject | GTP-BINDING PROTEIN | |
dc.subject | INSIGHTS | |
dc.subject | Life Sciences & Biomedicine - Other Topics | |
dc.subject | SIGNAL RECOGNITION PARTICLE | |
dc.subject | TRANSFER-RNA MODIFICATION | |
dc.title | Kissing G Domains of MnmE Monitored by X-Ray Crystallography and Pulse Electron Paramagnetic Resonance Spectroscopy | |
dc.type | journal article | |
dc.identifier.doi | 10.1371/journal.pbio.1000212 | |
dc.identifier.isi | ISI:000272031800002 | |
dc.description.volume | 7 | |
dc.description.issue | 10 | |
dc.contributor.orcid | 0000-0002-5888-0157 | |
dc.contributor.orcid | 0000-0002-5761-5968 | |
dc.contributor.researcherid | H-3791-2014 | |
dc.contributor.researcherid | C-1428-2009 | |
dc.publisher.place | 185 BERRY ST, STE 1300, SAN FRANCISCO, CA 94107 USA | |
dcterms.isPartOf.abbreviation | PLoS. Biol. | |
dcterms.oaStatus | Green Submitted, Green Published, gold | |
crisitem.author.dept | FB 04 - Physik | - |
crisitem.author.deptid | fb04 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | StHe633 | - |