DC Element | Wert | Sprache |
dc.contributor.author | Bhagawati, Maniraj | |
dc.contributor.author | Hoffmann, Simon | |
dc.contributor.author | Hoeffgen, Katharina S. | |
dc.contributor.author | Piehler, Jacob | |
dc.contributor.author | Busch, Karin B. | |
dc.contributor.author | Mootz, Henning D. | |
dc.date.accessioned | 2021-12-23T16:09:16Z | - |
dc.date.available | 2021-12-23T16:09:16Z | - |
dc.date.issued | 2020 | |
dc.identifier.issn | 14337851 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/8703 | - |
dc.description.abstract | Protein semi-synthesis inside live cells from exogenous and endogenous parts offers unique possibilities for studying proteins in their native context. Split-intein-mediated protein trans-splicing is predestined for such endeavors and has seen some successes, but a much larger variety of established split inteins and associated protocols is urgently needed. We characterized the association and splicing parameters of the Gp41-1 split intein, which favorably revealed a nanomolar affinity between the intein fragments combined with the exceptionally fast splicing rate. Following bead-loading of a chemically modified intein fragment precursor into live mammalian cells, we fluorescently labeled target proteins on their N- and C-termini with short peptide tags, thus ensuring minimal perturbation of their structure and function. In combination with a nuclear-entrapment strategy to minimize cytosolic fluorescence background, we applied our technique for super-resolution imaging and single-particle tracking of the outer mitochondrial protein Tom20 in HeLa cells. | |
dc.description.sponsorship | DFGGerman Research Foundation (DFG)European Commission [SPP1623, MO1073/5-2]; Alexander von Humboldt FoundationAlexander von Humboldt Foundation; Projekt DEAL; DFG (Cluster-of-excellence EXC1003 `Cells in Motion')German Research Foundation (DFG) [2017-07]; We gratefully acknowledge financial support by the DFG (SPP1623, MO1073/5-2 to H.D.M. and Cluster-of-excellence EXC1003 `Cells in Motion', FF project 2017-07 to K.B.B. and H.D.M.) and the Alexander von Humboldt Foundation (postdoctoral fellowship to M.B.). We thank Timo Dellmann for help with SPT data acquisition and the iBiOs Facility (PI 405/14-1) for help with dSTORM data acquisition and analysis (Dr. Rainer Kurre), and Christian P. Richter for developing softwares for analysis of SPT data. Open access funding enabled and organized by Projekt DEAL. | |
dc.language.iso | en | |
dc.publisher | WILEY-V C H VERLAG GMBH | |
dc.relation.ispartof | ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | |
dc.subject | Chemistry | |
dc.subject | Chemistry, Multidisciplinary | |
dc.subject | DNAE INTEIN | |
dc.subject | dSTORM | |
dc.subject | EVOLUTION | |
dc.subject | FLUOROPHORES | |
dc.subject | GENETIC-CODE | |
dc.subject | HIGHLY EFFICIENT | |
dc.subject | LABELING IN-VITRO | |
dc.subject | LIGATION | |
dc.subject | protein splicing | |
dc.subject | protein transduction | |
dc.subject | single-molecule studies | |
dc.subject | split intein | |
dc.subject | TAG | |
dc.title | In Cellulo Protein Semi-Synthesis from Endogenous and Exogenous Fragments Using the Ultra-Fast Split Gp41-1 Intein | |
dc.type | journal article | |
dc.identifier.doi | 10.1002/anie.202006822 | |
dc.identifier.isi | ISI:000568693800001 | |
dc.description.volume | 59 | |
dc.description.issue | 47 | |
dc.description.startpage | 21007 | |
dc.description.endpage | 21015 | |
dc.contributor.orcid | 0000-0003-0525-0191 | |
dc.contributor.researcherid | AAM-8374-2021 | |
dc.contributor.researcherid | ABH-8594-2020 | |
dc.identifier.eissn | 15213773 | |
dc.publisher.place | POSTFACH 101161, 69451 WEINHEIM, GERMANY | |
dcterms.isPartOf.abbreviation | Angew. Chem.-Int. Edit. | |
dcterms.oaStatus | Green Published, hybrid | |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | PiJa938 | - |