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Data for The Association of Defensin HNP-2 with Negatively Charged Membranes: A Combined Fluorescence and Linear Dichroism Study Open Access

Abstract from article: The association of defensin HNP-2 with negatively charged membranes has been studied using a new approach that combines fluorescence and linear dichroism (LD) spectroscopies with simulated LD spectra in order to characterise the binding kinetics and bound configurations of the peptide. Binding to membranes composed of mixtures of diacylglycerophosphocholines (PC) with either diacylglycerophosphoglycerol (PG) or diacylglycerophosphoserine (PS) was conducted at lipid:peptide ratios that yielded binding, but not membrane fusion. HNP-2 association with membranes under these conditions was a 2 stage-process, with both stages exhibiting first order kinetics. The fast initial step, with a half-life of < 1 min, was followed by a slower step with a half-life of > 3 min. Conversion between the states was estimated to have an enthalpy of activation of approximately 10 kJ/mol and an entropy of activation of − 0.2 kJ/K/mol. LD spectra corresponding to each of the membrane bound states were generated by non-linear regression using a standard kinetics model. These spectra are interpreted in comparison with spectra calculated using the program Dichrocalc and reveal that the peptide associates with membranes in a small number of stable configurations. All of these configurations have a significant proportion of β-sheet structure residing in the plane of the membrane. Two configurations support structures previously proposed for defensins in membranes.


Resource type
Creator: Sanderson, John 1
Creator: Pridmore, Catherine 1
Creator: Rodger, Alison 2
Contact person: Sanderson, John 1
1 Durham University, UK
2 University of Warwick, UK
Engineering and Physical Sciences Research Council
Research methods
Fluorescence spectroscopy; Linear Dichroism spectroscopy; calculations (Dichrocalc, APBS); Förster resonance energy transfer; dynamic light scattering; differential scanning calorimetry; circular dichroism spectroscopy.
Other description
Data generated during a study of defensin HNP-2 association with lipid membranes. Data are grouped by experiment. Information on files is located in each experiment directory, and in some cases where appropriate, in individual directories and files. Data types include: fluorescence (Figs. 2, 4, 6, S1); Linear Dichroism (Fig. 5, S8); Dichrocalc (Fig. 10, 11); APBS (Fig. 12, S17); FRET (Fig. S2, S3); DLS (Fig. S4, S5); DSC (Fig. S6); Circular Dichroism (Fig. S7).
Cited in
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA)

Durham University
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J.M. Sanderson
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17 May 2016, 14:05:35
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