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. 2018 Dec 22;47(4):1935–1949. doi: 10.1093/nar/gky1280

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 2. — Active distortion of the HJ by GEN1 (A) Inline-label schematic of the HJ structural distortion by GEN1 based on the proposed model (36). (B) FRET histogram of inline-label X₀ showing a single peak (E ≈ 0.40). At saturating GEN1 concentration, the major low FRET peak (E ≈ 0.25) is attributed to GEN1-Iso(1,3) while the minor high FRET peak (E ≈ 0.46) results from GEN1-Iso(2,4). (C) FRET histogram of J₇ showing a single peak (E ≈ 0.48). GEN1 binding to Iso(1,3) and Iso(2,4) yields two nearly equal populations of low and high FRET isomers, respectively. (D) Upper panel: FRET time trace of inline-label X₀. Lower panel: GEN1 is flown in Ca²⁺ buffer to detect the onset of binding. A stable complex persists throughout the acquisition time. (E) GEN1 binding to adjacent-label Iso(1,3) and Iso(2,4) forms a low FRET structure (E ≈ 0.32). (F) FRET histogram of X₀ exhibits a major high FRET peak (E ≈ 0.60) corresponding to Iso(1,3) and a lower FRET peak (E ≈ 0.40) for Iso(2,4). At saturating GEN1 concentration, the whole histogram is transformed into a single low FRET peak. (G) FRET histogram of J₇ exhibits two nearly equal populations at high and low FRET corresponding to Iso(1,3) and Iso(2,4), respectively. Binding of GEN1 forms a low FRET structure corresponding to a single peak (E ≈ 0.37). (H) Upper panel: FRET time trace of adjacent-label X₀. Lower panel: GEN1 binding forms a stable low FRET state (E ≈ 0.32). The uncertainties in panels B, C, F, G represent the standard deviations from two or more experiments.