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FRET-FCS Detection of Intra-Lobe Dynamics in Calmodulin
dc.contributor.author | Price, E. Shane | |
dc.contributor.author | Aleksiejew, Marek | |
dc.contributor.author | Johnson, Carey K. | |
dc.date.accessioned | 2017-04-27T19:15:54Z | |
dc.date.available | 2017-04-27T19:15:54Z | |
dc.date.issued | 2011-07-28 | |
dc.identifier.citation | Price, E. S., Aleksiejew, M., & Johnson, C. K. (2011). FRET-FCS Detection of Intra-Lobe Dynamics in Calmodulin. The Journal of Physical Chemistry. B, 115(29), 9320–9326. http://doi.org/10.1021/jp203743m | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/23847 | |
dc.description.abstract | Fluorescence correlation spectroscopy (FCS) can be coupled with Förster resonance energy transfer (FRET) to detect intramolecular dynamics of proteins on the microsecond time scale. Here we describe application of FRET-FCS to detect fluctuations within the N-terminal and C-terminal domains of the Ca2+-signaling protein calmodulin. Intramolecular fluctuations were resolved byglobal fitting of the two fluorescence autocorrelation functions (green-green and red-red) together with the two cross-correlation functions (green-red and red-green). To match the Förster radius forFRET to the dimensions of the N-terminal and C-terminal domains, a near-infrared acceptor fluorophore (Atto 740) was coupled with a green-emitting donor (Alexa Fluor 488). Fluctuations were detected in both domains on the time scale of 30 to 40 μs. In the N-terminal domain, the amplitude of the fluctuations was dependent on occupancy of Ca2+ binding sites. A high amplitude of dynamics in apo-calmodulin (in the absence of Ca2+) was nearly abolished at a high Ca2+ concentration. For the C-terminal domain the dynamic amplitude changed little with Ca2+ concentration. The Ca2+ dependence of dynamics for the N-terminal domain suggests that the fluctuations detected by FCS in the N-terminal domain are coupled to the opening and closing of the EF-hand Ca2+-binding loops. | en_US |
dc.publisher | American Chemical Society | en_US |
dc.rights | This document is the Accepted Manuscript version of a Published Work that appeared in final form in the Journal of Physical Chemistry B, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://dx.doi.org/10.1021/jp203743m. | en_US |
dc.subject | Calmodulin | en_US |
dc.subject | Fluorescence correlation spectroscopy | en_US |
dc.subject | FRET | en_US |
dc.subject | Protein dynamics | en_US |
dc.title | FRET-FCS Detection of Intra-Lobe Dynamics in Calmodulin | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Price, E. Shane | |
kusw.kuauthor | Marek, Aleksiejaw | |
kusw.kuauthor | Johnson, Carey K. | |
kusw.kudepartment | Chemistry | en_US |
dc.identifier.doi | 10.1021/jp203743m | en_US |
kusw.oaversion | Scholarly/refereed, author accepted manuscript | en_US |
kusw.oapolicy | This item meets KU Open Access policy criteria. | en_US |
dc.identifier.pmid | PMC3151148 | en_US |
dc.rights.accessrights | openAccess |