Abstract
Purpose. The weak aqueous interaction between the protein lactate dehydrogenase (LDH) and the nonionic surfactant Tween 80 has been investigated, because weak protein-amphiphile interactions are of significant importance in pharmaceutical formulations, but are experimentally hard to determine. The system LDH/sodium dodecyl sulphate (SDS) was used as reference because SDS, by its strong protein binding, denatures LDH completely.
Methods. Fluorescence spectroscopy with pyrene and 1,3-bis(1-phenyl)propane (P3P) as probes, intrinsic protein fluorescence and NMR spectroscopy have been used.
Results. The fluorescence probe pyrene monitors a weak Tween-LDH interaction, detectable below the critical micelle concentration of ordinary Tween micelles. The microviscosity probe P3P shows a surfactant-induced denaturation in the case of LDH/SDS but not in the case of LDH/Tween 80. Intrinsic LDH fluorescence verifies this behavior. Pulsed-gradient spin-echo NMR was also used to verify the weak LDH-Tween 80 interaction.
Conclusions. A weak interaction between LDH and Tween 80 occurs at hydrophobic zones of the protein, but it is not strong enough to denature LDH. The experimental outline used here provides a useful approach for mapping the very weak protein-amphiphile interactions often present in pharmaceutical formulations.
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Hillgren, A., Evertsson, H. & Aldén, M. Interaction Between Lactate Dehydrogenase and Tween 80 in Aqueous Solution. Pharm Res 19, 504–510 (2002). https://doi.org/10.1023/A:1015156031381
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DOI: https://doi.org/10.1023/A:1015156031381