Interaction between lactate dehydrogenase and Tween 80 in aqueous solution.
OBJECTIVE
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(lphenyl)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.
Hillgren
Anna
A
Department of Pharmacy, Uppsala University, Uppsala Biomedical Center, Sweden.
Evertsson
Hans
H
Aldén
Maggie
M
D000818:Animals; D002152:Calorimetry, Differential Scanning / Q000379:methods; D004347:Drug Interactions; D007770:L-Lactate Dehydrogenase / Q000737:chemistry* / Q000493:pharmacokinetics; D019999:Pharmaceutical Solutions / Q000737:chemistry / Q000493:pharmacokinetics; D011136:Polysorbates / Q000737:chemistry* / Q000493:pharmacokinetics; D011721:Pyrenes / Q000493:pharmacokinetics; D011817:Rabbits; D013050:Spectrometry, Fluorescence / Q000379:methods; D014783:Viscosity; D014867:Water / Q000737:chemistry*
D019999:Pharmaceutical Solutions; D011136:Polysorbates; D011721:Pyrenes; D014867:Water; C030984:pyrene; D007770:L-Lactate Dehydrogenase