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Detecting sequential bond formation using three-dimensional thermal fluctuation analysis
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SYSNO ASEP 0333759 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Detecting sequential bond formation using three-dimensional thermal fluctuation analysis Author(s) Bartsch, T. (US)
Fisinger, S. (US)
Kochanczyk, M.D. (US)
Huang, R. (US)
Jonáš, Alexandr (UPT-D) RID, SAI, ORCID
Florin, E. L. (DE)Source Title ChemPhysChem. - : Wiley - ISSN 1439-4235
Roč. 10, 9-10 (2009), s. 1541-1547Number of pages 7 s. Language eng - English Country DE - Germany Keywords biophysics ; mechanical properties ; optical tweezers ; single-molecule studies ; thermal fluctuations Subject RIV BO - Biophysics CEZ AV0Z20650511 - UPT-D (2005-2011) UT WOS 000267928100028 DOI 10.1002/cphc.200900211 Annotation We present a novel experimental method that solves two key problems in nondestructive mechanical studies of small biomolecules at the single-molecule level, namely the confirmation of single-molecule conditions and the discrimination against nonspecific binding. A biotin avidin ligand-receptor couple is spanned between a glass slide and a 1 mu m latex particles using short linker molecules. Optical tweezers are used to initiate bond formation and to follow the particles thermal position fluctuations with nanometer spatial and microsecond temporal resolution. Here we show that each step in the specific binding process leads to an abrupt change in the magnitude of the particle's thermal position fluctuations, allowing us to count the number of bonds formed one by one. Moreover, three-dimensional position histograms calculated from the particle's fluctuations can be separated into well-defined categories reflecting different binding conditions. Workplace Institute of Scientific Instruments Contact Martina Šillerová, sillerova@ISIBrno.Cz, Tel.: 541 514 178 Year of Publishing 2010
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