Xenbase Image ID: 123435

FIGURE 1. Fluorescence change as a function of protein binding. A, fluorescence (A.U.) as a function of wavelength (nm), with closed circles/squares representing the signal from the excitation of Alexa-488 (yNap1m*) in the presence (closed squares) or absence (closed circles) of H2A/H2B and the open circles/squares representing Alexa-546 (H2A/H2B(T112C)) in the presence (open squares) or absence (open circles) of yNap1. B, normalized fluorescence change as a function of either H2A/H2B titrated into Alexa-546-labeled yNap1 (closed squares) or yNap1 titrated into Alexa-546 labeled H2A/H2B (open circles). C, change in fluorescence plotted as a function of the ratio of H2A/H2B to yNap1. The intersection of the linear phase with the plateau is equal to the molar ratio at which yNap1 is saturated. These data suggest that two H2A/H2B dimers bind one yNap1 dimer or that one H2A/H2B binds one yNap1 monomer. The conditions were 20–50 mm Tris, pH 7.5, 1 mm dithiothreitol, 0.1 mg/ml bovine serum albumin, 90–120 mm NaCl, (1–5) × 10–10 m Alexa-labeled protein, and (0–5) × 10–7 m non-labeled protein. For results of the fit see Table 1. Image published in: Andrews AJ et al. (2008) Copyright © 2008, The American Society for Biochemistry and Molecular Biology, Inc. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

Return to previous page