The association of defensin HNP-2 with negatively charged membranes: A combined fluorescence and linear dichroism study

Biochimica Et Biophysica Acta
Catherine J PridmoreJohn M Sanderson

Abstract

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(-1) and an entropy of activation of -0.2 kJ K mol(-1). LD spectra corresponding to each of the membrane bound states were generated by non-linear regression using a standard kinetic 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 co...Continue Reading

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Citations

May 19, 2017·FEMS Microbiology Reviews·María Eugenia Pachón-IbáñezJavier Sánchez-Céspedes
Aug 26, 2017·Biophysics Reviews·John M Sanderson, Frances Separovic

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