Four-color single-molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes
DC Element | Wert | Sprache |
---|---|---|
dc.contributor.author | DeRocco, Vanessa C. | |
dc.contributor.author | Anderson, Trevor | |
dc.contributor.author | Piehler, Jacob | |
dc.contributor.author | Erie, Dorothy A. | |
dc.contributor.author | Weninger, Keith | |
dc.date.accessioned | 2021-12-23T16:17:51Z | - |
dc.date.available | 2021-12-23T16:17:51Z | - |
dc.date.issued | 2010 | |
dc.identifier.issn | 07366205 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/12437 | - |
dc.description.abstract | To enable studies of conformational changes within multimolecular complexes, we present a simultaneous, four-color single molecule fluorescence methodology implemented with total internal reflection illumination and camera-based, wide-field detection. We further demonstrate labeling histidine-tagged proteins noncovalently with Tris-nitrilotriacetic acid (Tris-NTA)-conjugated dyes to achieve single molecule detection. We combine these methods to colocalize the mismatch repair protein MutS alpha. on DNA while monitoring MutS alpha-induced DNA bending using Forster resonance energy transfer (FRET) and to monitor assembly of membrane-tethered SNARE protein complexes. | |
dc.description.sponsorship | National Institutes of Health (NIH)United States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [R01 GM079480, GM080294]; American Cancer SocietyAmerican Cancer Society [RSG-03-047, RSG-10-048]; Career Award at the Scientific Interface from the Burroughs Wellcome FundBurroughs Wellcome Fund; National Institute of General Medical SciencesUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [F31GM087096]; NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCESUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [R01GM079480, F31GM087096, R01GM080294] Funding Source: NIH RePORTER; We thank Susan Doyle of the Erie Lab and Alan Clark of the Kunkel Lab for providing purified yMutS alpha. This work was supported in part by the National Institutes of Health (NIH; grant nos. R01 GM079480 and GM080294) and the American Cancer Society [grant nos. RSG-03-047 (to D.A.E.) and RSG-10-048 (to K.W.)]. The research of K.W. is supported in part by a Career Award at the Scientific Interface from the Burroughs Wellcome Fund. V.C.D. was also supported in part by the National Institute of General Medical Sciences (award no. F31GM087096). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences or the NIH. This paper is subject to the NIH Public Access Policy. | |
dc.language.iso | en | |
dc.publisher | FUTURE SCI LTD | |
dc.relation.ispartof | BIOTECHNIQUES | |
dc.subject | ALTERNATING-LASER EXCITATION | |
dc.subject | Biochemical Research Methods | |
dc.subject | Biochemistry & Molecular Biology | |
dc.subject | DNA mismatch repair | |
dc.subject | FRET | |
dc.subject | FUSION | |
dc.subject | HISTIDINE-TAGGED PROTEINS | |
dc.subject | MECHANISM | |
dc.subject | Msh2 | |
dc.subject | Msh6 | |
dc.subject | Munc-18 | |
dc.subject | MutS | |
dc.subject | REAL-TIME | |
dc.subject | RECOGNITION | |
dc.subject | RESONANCE ENERGY-TRANSFER | |
dc.subject | SNARE COMPLEX | |
dc.subject | SNARE proteins | |
dc.subject | SPECTROSCOPY | |
dc.title | Four-color single-molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes | |
dc.type | journal article | |
dc.identifier.doi | 10.2144/000113551 | |
dc.identifier.isi | ISI:000287719300012 | |
dc.description.volume | 49 | |
dc.description.issue | 5 | |
dc.description.startpage | 807+ | |
dc.identifier.eissn | 19409818 | |
dc.publisher.place | UNITED HOUSE, 2 ALBERT PL, LONDON, N3 1QB, ENGLAND | |
dcterms.isPartOf.abbreviation | Biotechniques | |
dcterms.oaStatus | Green Accepted, Green Submitted, gold | |
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 | - |
Seitenaufrufe
4
Letzte Woche
0
0
Letzter Monat
0
0
geprüft am 01.05.2024