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. Author manuscript; available in PMC: 2009 Sep 19.
Published in final edited form as: ACS Chem Biol. 2008 Aug 22;3(9):555–566. doi: 10.1021/cb800059u

Figure 3.

Figure 3

Technical aspects of dynamic fluorescence depolarization data collection. a) Representative physical model (3-component) for multiple independent molecular motions and matching equations applicable to rotational correlation times (τi) differing by at least one order of magnitude. The BODIPY-FL fluorophore is shown in green. Fi is the fractional amplitude for the anisotropy decay corresponding to each of the motions. In the case of 2-component RNC dynamics, the intermediate timescale (I) term is lost. b) Representative experimental fluorescence anisotropy data for short (apoMb16) and long (apoMb153) ribosome-bound nascent chains and associated 2- and 3-component fits, respectively. c) Multiexponential curve fitting (2- and 3-component analysis) and residuals for ribosome-bound ApoMb16. d) ApoMb153 analyzed similarly to panel c and 2-component fit with variable r0. e) Simulation illustrating the unique features of typical 2-and 3-component frequency-domain anisotropy decays. f) Expected phase changes upon varying the fraction of intermediate (fI) and fast timescale motions, to illustrate the progression from 2-(fI = 0) to 3-component decays. The 2- and 3-component phase change plots of panel e coincide with simulations A and C. g) Parameters used for the simulations in panel f.