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Spectroscopic and calorimetric studies on the interaction of human serum albumin with DPPC/PEG:2000-DPPE membranes

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Abstract

Site specific spectroscopic techniques and differential scanning calorimetry were used to study human serum albumin (HSA) in the absence and in the presence of membranes composed of dipalmitoylphosphatidylcholine (DPPC) and poly(ethylene glycol:2000)-dipalmitoylphosphatidylethanolamine (PEG:2000-DPPE). Electron spin resonance (ESR) of a maleimide spin-label (5-MSL) covalently bound to the free sulfhydryl group at the unique cystein Cys-34 in domain I, intrinsic fluorescence of the single tryptophan Trp-214 in domain II, and extrinsic fluorescence of p-nitrophenyl anthranilate conjugated with tyrosine Tyr-411 in domain III were employed to study HSA dispersions with or without polymer-grafted membranes. On adsorbing at the DPPC membrane surfaces, domain I assumes a more loosened conformation and partitioning of the spin-labelled protein between the aqueous phase and the interfacial region of lipid membranes is observed by ESR. Domain II and III undergo a local structural arrangement which leads Trp-214 and Tyr-411 to come closer and causes intrinsic fluorescence quenching. The influence of DPPC bilayers on HSA is characterized both by a decrease of the thermal unfolding enthalpy and by a slight increase of the transition temperature, T t, of the protein. The lipid induced effects on HSA are progressively reduced on increasing the amounts of PEG:2000-DPPE mixed with DPPC from the mushroom regime to the brush regime. Primary protein adsorption at the lipid surfaces is abolished at 1 mol% of the polymer-lipid, whereas the secondary protein adsorption at the polymer-brush leads to a further increase of both transition enthalpy and T t relative to the case of aqueous dispersions of HSA alone.

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Acknowledgments

Manuela Pantusa thanks the University of Calabria for a research grant. This work was financially supported by University of Calabria.

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  1. Dipartimento di Fisica, Laboratorio di Biofisica Molecolare and UdR CNISM, Università della Calabria, 87036, Arcavacata di Rende (CS), Italy

    Manuela Pantusa, Luigi Sportelli & Rosa Bartucci

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  1. Manuela Pantusa
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  2. Luigi Sportelli
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  3. Rosa Bartucci
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Correspondence to Rosa Bartucci.

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Pantusa, M., Sportelli, L. & Bartucci, R. Spectroscopic and calorimetric studies on the interaction of human serum albumin with DPPC/PEG:2000-DPPE membranes. Eur Biophys J 37, 961–973 (2008). https://doi.org/10.1007/s00249-008-0314-z

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  • DOI: https://doi.org/10.1007/s00249-008-0314-z

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