期刊
CHEMICAL PHYSICS
卷 422, 期 -, 页码 238-245出版社
ELSEVIER
DOI: 10.1016/j.chemphys.2012.11.018
关键词
Calmodulin; FRET; Single-molecule fluorescence; Fluorescence dyes; Bayesian analysis; Nested sampling; Maximum entropy; Calcium/calmodulin dependent protein kinase II
资金
- National Science Foundation [CHE-0710515]
- NIH [GM08545]
- STFC [ST/J00152X/1] Funding Source: UKRI
- Science and Technology Facilities Council [ST/J00152X/1] Funding Source: researchfish
We analyzed single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest compares favorably with maximum entropy analysis for simulated data, judged by the Bayesian evidence. FRET efficiency distributions recovered for calmodulin labeled with two different FRET dye pairs depended on the dye pair and changed upon Ca2+ binding. We also looked at the FRET efficiency distributions of calmodulin bound to the calcium/calmodulin dependent protein kinase II (CaMKII) binding domain. For both dye pairs, the FRET efficiency distribution collapsed to a single peak in the case of calmodulin bound to the CaMKII peptide. These measurements strongly suggest that consideration of dye-protein interactions is crucial in forming an accurate picture of protein conformations from FRET data. (C) 2012 Elsevier B.V. All rights reserved.
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