Holtkamp, Wolf

[["dc.bibliographiccitation.firstpage","1104"],["dc.bibliographiccitation.issue","6264"],["dc.bibliographiccitation.journal","Science"],["dc.bibliographiccitation.lastpage","1107"],["dc.bibliographiccitation.volume","350"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Kokic, Goran"],["dc.contributor.author","Jäger, Marcus"],["dc.contributor.author","Mittelstaet, Joerg"],["dc.contributor.author","Komar, Anton A."],["dc.contributor.author","Rodnina, Marina V."],["dc.date.accessioned","2017-09-07T11:54:52Z"],["dc.date.available","2017-09-07T11:54:52Z"],["dc.date.issued","2015"],["dc.description.abstract","Protein domains can fold into stable tertiary structures while they are synthesized on the ribosome. We used a high-performance, reconstituted in vitro translation system to investigate the folding of a small five-helix protein domain-the N-terminal domain of Escherichia coli N5-glutamine methyltransferase HemK-in real time. Our observations show that cotranslational folding of the protein, which folds autonomously and rapidly in solution, proceeds through a compact, non-native conformation that forms within the peptide tunnel of the ribosome. The compact state rearranges into a native-like structure immediately after the full domain sequence has emerged from the ribosome. Both folding transitions are rate-limited by translation, allowing for quasi-equilibrium sampling of the conformational space restricted by the ribosome. Cotranslational folding may be typical of small, intrinsically rapidly folding protein domains."],["dc.identifier.doi","10.1126/science.aad0344"],["dc.identifier.gro","3141785"],["dc.identifier.isi","000366422600047"],["dc.identifier.pmid","26612953"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1046"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1095-9203"],["dc.relation.issn","0036-8075"],["dc.title","Cotranslational protein folding on the ribosome monitored in real time"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","e130"],["dc.bibliographiccitation.issue","22"],["dc.bibliographiccitation.journal","Nucleic Acids Research"],["dc.bibliographiccitation.lastpage","e130"],["dc.bibliographiccitation.volume","46"],["dc.contributor.author","Haase, Nadin"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Lipowsky, Reinhard"],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Rudorf, Sophia"],["dc.date.accessioned","2021-06-01T10:51:20Z"],["dc.date.available","2021-06-01T10:51:20Z"],["dc.date.issued","2018"],["dc.identifier.doi","10.1093/nar/gky740"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/86981"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-425"],["dc.relation.eissn","1362-4962"],["dc.relation.issn","0305-1048"],["dc.title","Decomposition of time-dependent fluorescence signals reveals codon-specific kinetics of protein synthesis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1073"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","The EMBO journal"],["dc.bibliographiccitation.lastpage","1085"],["dc.bibliographiccitation.volume","33"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Cunha, Carlos E. da"],["dc.contributor.author","Peske, Frank"],["dc.contributor.author","Konevega, Andrey L."],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.contributor.author","Rodnina, Marina V."],["dc.date.accessioned","2018-01-26T06:04:53Z"],["dc.date.available","2018-01-26T06:04:53Z"],["dc.date.issued","2014"],["dc.description.abstract","Elongation factor G (EF-G) promotes the movement of two tRNAs and the mRNA through the ribosome in each cycle of peptide elongation. During translocation, the tRNAs transiently occupy intermediate positions on both small (30S) and large (50S) ribosomal subunits. How EF-G and GTP hydrolysis control these movements is still unclear. We used fluorescence labels that specifically monitor movements on either 30S or 50S subunits in combination with EF-G mutants and translocation-specific antibiotics to investigate timing and energetics of translocation. We show that EF-G-GTP facilitates synchronous movements of peptidyl-tRNA on the two subunits into an early post-translocation state, which resembles a chimeric state identified by structural studies. EF-G binding without GTP hydrolysis promotes only partial tRNA movement on the 50S subunit. However, rapid 30S translocation and the concomitant completion of 50S translocation require GTP hydrolysis and a functional domain 4 of EF-G. Our results reveal two distinct modes for utilizing the energy of EF-G binding and GTP hydrolysis and suggest that coupling of GTP hydrolysis to translocation is mediated through rearrangements of the 30S subunit."],["dc.identifier.doi","10.1002/embj.201387465"],["dc.identifier.pmid","24614227"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11834"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.relation.eissn","1460-2075"],["dc.title","GTP hydrolysis by EF-G synchronizes tRNA movement on small and large ribosomal subunits"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","908"],["dc.bibliographiccitation.issue","10"],["dc.bibliographiccitation.journal","BioEssays"],["dc.bibliographiccitation.lastpage","918"],["dc.bibliographiccitation.volume","36"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.contributor.author","Rodnina, Marina V."],["dc.date.accessioned","2017-09-07T11:45:30Z"],["dc.date.available","2017-09-07T11:45:30Z"],["dc.date.issued","2014"],["dc.description.abstract","The translocation of tRNAs through the ribosome proceeds through numerous small steps in which tRNAs gradually shift their positions on the small and large ribosomal subunits. The most urgent questions are: (i) whether these intermediates are important; (ii) how the ribosomal translocase, the GTPase elongation factor G (EF-G), promotes directed movement; and (iii) how the energy of GTP hydrolysis is coupled to movement. In the light of recent advances in biophysical and structural studies, we argue that intermediate states of translocation are snapshots of dynamic fluctuations that guide the movement. In contrast to current models of stepwise translocation, kinetic evidence shows that the tRNAs move synchronously on the two ribosomal subunits in a rapid reaction orchestrated by EF-G and GTP hydrolysis. EF-G combines the energy regimes of a GTPase and a motor protein and facilitates tRNA movement by a combination of directed Brownian ratchet and power stroke mechanisms."],["dc.identifier.doi","10.1002/bies.201400076"],["dc.identifier.gro","3142042"],["dc.identifier.isi","000342914600002"],["dc.identifier.pmid","25118068"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3901"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1521-1878"],["dc.relation.issn","0265-9247"],["dc.title","Synchronous tRNA movements during translocation on the ribosome are orchestrated by elongation factor G and GTP hydrolysis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","83"],["dc.bibliographiccitation.journal","Current Opinion in Structural Biology"],["dc.bibliographiccitation.lastpage","89"],["dc.bibliographiccitation.volume","42"],["dc.contributor.author","Thommen, Michael"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Rodnina, Marina V."],["dc.date.accessioned","2018-01-17T12:55:22Z"],["dc.date.available","2018-01-17T12:55:22Z"],["dc.date.issued","2017"],["dc.description.abstract","Proteins are synthesized as linear polymers and have to fold into their native structure to fulfil various functions in the cell. Folding can start co-translationally when the emerging peptide is still attached to the ribosome and is guided by the environment of the polypeptide exit tunnel and the kinetics of translation. Major questions are: When does co-translational folding begin? What is the role of the ribosome in guiding the nascent peptide towards its native structure? How does translation elongation kinetics modulate protein folding? Here we suggest how novel structural and biophysical approaches can help to probe the interplay between the ribosome and the emerging peptide and present future challenges in understanding co-translational folding."],["dc.identifier.doi","10.1016/j.sbi.2016.11.020"],["dc.identifier.pmid","27940242"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11695"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.relation.eissn","1879-033X"],["dc.title","Co-translational protein folding: progress and methods"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","221"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Zeitschrift für Gastroenterologie"],["dc.bibliographiccitation.lastpage","227"],["dc.bibliographiccitation.volume","38"],["dc.contributor.author","Holtkamp, W."],["dc.contributor.author","Mueller, Walter J."],["dc.date.accessioned","2018-11-07T09:47:06Z"],["dc.date.available","2018-11-07T09:47:06Z"],["dc.date.issued","2000"],["dc.description.abstract","Percutaneous radiofrequency ablation (HFTT) is a new therapeutic technique for the treatment of inoperable primary and secondary liver tumors. We report our initial experience using a newly developed perfusion electrode. Twelve liver tumors (11 metastases of colorectal tumors, 1 hepatocellular carcinoma) were treated in 5 inoperable patients. The patients had 1 to 3 liver tumors. All lesions were cytologically confirmed and measured 12-47 mm. The technique was approved by the institutional review board and informend consent was obtained from all patients. A 12-mm-needle electrode with a 15-mm-active tip was introduced into the liver tumors under ultrasound guidance and tumors were coagulated with radiofrequency energy of 350 kHz. The needle electrodes were perfused with 0.9% saline during coagulation to increase the volume of coagulation necrosis without tissue vaporization. The serial changes in tumor size after therapy were evaluated with spiral CT imaging. Dynamic CT showed that unenhanced areas indicative of coagulation necrosis developed in all tumors. In 8 of 12 tumors no signs of recurrence ap peared during the observation period of 7 (5-12) months. No major complications were observed. Our preliminary experience suggests that percutaneous radiofrequency coagulation can be a simple, safe and potentially effective treatment for selected patients with inoperable liver tumors. The results justify further studies to investigate the possible role in clinical practice."],["dc.identifier.doi","10.1055/s-2000-14861"],["dc.identifier.isi","000087200200002"],["dc.identifier.pmid","10768244"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/35034"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Demeter Verlag Georg Thieme Verlag"],["dc.relation.issn","0044-2771"],["dc.title","Percutaneous ultrasound-guided high frequency thermal therapy [HFTT] of hepatic tumors with perfused fine-needle electrodes"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","12186"],["dc.bibliographiccitation.issue","22"],["dc.bibliographiccitation.journal","Nucleic Acids Research"],["dc.bibliographiccitation.lastpage","12187"],["dc.bibliographiccitation.volume","46"],["dc.contributor.author","Haase, Nadin"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Lipowsky, Reinhard"],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Rudorf, Sophia"],["dc.date.accessioned","2021-06-01T10:51:20Z"],["dc.date.available","2021-06-01T10:51:20Z"],["dc.date.issued","2018"],["dc.identifier.doi","10.1093/nar/gky1101"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/86980"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-425"],["dc.relation.eissn","1362-4962"],["dc.relation.issn","0305-1048"],["dc.title","Decomposition of time-dependent fluorescence signals reveals codon-specific kinetics of protein synthesis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","e24315"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Translation"],["dc.bibliographiccitation.volume","1"],["dc.contributor.author","Cunha, Carlos E."],["dc.contributor.author","Belardinelli, Riccardo"],["dc.contributor.author","Peske, Frank"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.contributor.author","Rodnina, Marina V."],["dc.date.accessioned","2018-01-26T08:50:21Z"],["dc.date.available","2018-01-26T08:50:21Z"],["dc.date.issued","2013"],["dc.description.abstract","Elongation factor G (EF-G) is a GTPase that catalyzes tRNA and mRNA translocation during the elongation cycle of protein synthesis. The GTP-bound state of the factor on the ribosome has been studied mainly with non-hydrolyzable analogs of GTP, which led to controversial conclusions about the role of GTP hydrolysis in translocation. Here we describe a mutant of EF-G in which the catalytic His91 is replaced with Ala. The mutant EF-G does not hydrolyze GTP, but binds GTP with unchanged affinity, allowing us to study the function of the authentic GTP-bound form of EF-G in translocation. Utilizing fluorescent reporter groups attached to the tRNAs, mRNA, and the ribosome we compile the velocity map of translocation seen from different perspectives. The data suggest that GTP hydrolysis accelerates translocation up to 30-fold and facilitates conformational rearrangements of both 30S subunit (presumably the backward rotation of the 30S head) and EF-G that lead to the dissociation of the factor. Thus, EF-G combines the energy regime characteristic for motor proteins, accelerating movement by a conformational change induced by GTP hydrolysis, with that of a switch GTPase, which upon Pi release switches the conformations of EF-G and the ribosome to low affinity, allowing the dissociation of the factor."],["dc.format.extent","1-11"],["dc.identifier.doi","10.4161/trla.24315"],["dc.identifier.pmid","26824016"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11848"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.title","Dual use of GTP hydrolysis by elongation factor G on the ribosome"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","11858"],["dc.bibliographiccitation.issue","20"],["dc.bibliographiccitation.journal","Nucleic acids research"],["dc.bibliographiccitation.lastpage","11866"],["dc.bibliographiccitation.volume","45"],["dc.contributor.author","Mercier, Evan"],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Rodnina, Marina V."],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2018-01-17T12:49:58Z"],["dc.date.available","2018-01-17T12:49:58Z"],["dc.date.issued","2017-11-16"],["dc.description.abstract","The bacterial signal recognition particle (SRP) is part of the machinery that targets ribosomes synthesizing membrane proteins to membrane-embedded translocons co-translationally. Recognition of nascent membrane proteins occurs by virtue of a hydrophobic signal-anchor sequence (SAS) contained in the nascent chain, usually at the N terminus. Here we use fluorescence-based stopped-flow to monitor SRP-ribosome interactions with actively translating ribosomes while an SRP substrate is synthesized and emerges from the peptide exit tunnel. The kinetic analysis reveals that, at cellular concentrations of ribosomes and SRP, SRP rapidly binds to translating ribosomes prior to the emergence of an SAS and forms an initial complex that rapidly rearranges to a more stable engaged complex. When the growing peptide reaches a length of ∼50 amino acids and the SAS is partially exposed, SRP undergoes another conformational change which further stabilizes the complex and initiates targeting of the translating ribosome to the translocon. These results provide a reconciled view on the timing of high-affinity targeting complex formation, while emphasizing the existence of preceding SRP recruitment steps under conditions of ongoing translation."],["dc.identifier.doi","10.1093/nar/gkx888"],["dc.identifier.pmid","29149347"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11688"],["dc.identifier.url","https://sfb1190.med.uni-goettingen.de/production/literature/publications/14"],["dc.language.iso","en"],["dc.notes.status","zu prüfen"],["dc.relation","SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente"],["dc.relation","SFB 1190 | P16: Co-translationaler Einbau von Proteinen in die bakterielle Plasmamembran"],["dc.relation.eissn","1362-4962"],["dc.relation.workinggroup","RG Rodnina"],["dc.rights","CC BY-NC 4.0"],["dc.title","Signal recognition particle binds to translating ribosomes before emergence of a signal anchor sequence"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.subtype","original_ja"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","517"],["dc.bibliographiccitation.issue","7519"],["dc.bibliographiccitation.journal","Nature"],["dc.bibliographiccitation.lastpage","522"],["dc.bibliographiccitation.volume","513"],["dc.contributor.author","Loubresse, Nicolas Garreau de"],["dc.contributor.author","Prokhorova, Irina V."],["dc.contributor.author","Holtkamp, Wolf"],["dc.contributor.author","Rodnina, Marina V."],["dc.contributor.author","Yusupova, Gulnara"],["dc.contributor.author","Yusupov, Marat"],["dc.date.accessioned","2017-09-07T11:45:30Z"],["dc.date.available","2017-09-07T11:45:30Z"],["dc.date.issued","2014"],["dc.description.abstract","The ribosome is a molecular machine responsible for protein synthesis and a major target for small-molecule inhibitors. Compared to the wealth of structural information available on ribosome-targeting antibiotics in bacteria, our understanding of the binding mode of ribosome inhibitors in eukaryotes is currently limited. Here we used X-ray crystallography to determine 16 high-resolution structures of 80S ribosomes from Saccharomyces cerevisiae in complexes with 12 eukaryote-specific and 4 broad-spectrum inhibitors. All inhibitors were found associated with messenger RNA and transfer RNA binding sites. In combination with kinetic experiments, the structures suggest a model for the action of cycloheximide and lactimidomycin, which explains why lactimidomycin, the larger compound, specifically targets the first elongation cycle. The study defines common principles of targeting and resistance, provides insights into translation inhibitor mode of action and reveals the structural determinants responsible for species selectivity which could guide future drug development."],["dc.identifier.doi","10.1038/nature13737"],["dc.identifier.gro","3142047"],["dc.identifier.isi","000342075800033"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3956"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1476-4687"],["dc.relation.issn","0028-0836"],["dc.title","Structural basis for the inhibition of the eukaryotic ribosome"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]