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Exploring the pH-Induced Functional Phase Space of Human Serum Albumin by EPR Spectroscopy

1
Institute of Chemistry, Martin Luther University Halle-Wittenberg, Von-Danckelmann-Platz 4, 06120 Halle (Saale), Germany
2
Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Strasse 4, 06120 Halle (Saale), Germany
3
Fraunhofer Institute for Microstructure of Materials and Systems IMWS, Walter-Hülse-Strasse 1, 06120 Halle (Saale), Germany
*
Author to whom correspondence should be addressed.
Magnetochemistry 2018, 4(4), 47; https://doi.org/10.3390/magnetochemistry4040047
Received: 24 September 2018 / Revised: 25 October 2018 / Accepted: 26 October 2018 / Published: 1 November 2018
(This article belongs to the Special Issue Electron Paramagnetic Resonance)
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Abstract

A systematic study on the self-assembled solution system of human serum albumin (HSA) and paramagnetic doxyl stearic acid (5-DSA and 16-DSA) ligands is reported covering the broad pH range 0.7–12.9, mainly using electron paramagnetic resonance (EPR) methods. It is tested to which extent the pH-induced conformational isomers of HSA reveal themselves in continuous wave (CW) EPR spectra from this spin probing approach in comparison to an established spin-labeling strategy utilizing 3-maleimido proxyl (5-MSL). Most analyses are conducted on empirical levels with robust strategies that allow for the detection of dynamic changes of ligand, as well as protein. Special emphasis has been placed on the EPR spectroscopic detection of a molten globule (MG) state of HSA that is typically found by the fluorescent probe 8-Anilino- naphthalene-1-sulfonic acid (ANS). Moreover, four-pulse double electron-electron resonance (DEER) experiments are conducted and substantiated with dynamic light scattering (DLS) data to determine changes in the solution shape of HSA with pH. All results are ultimately combined in a detailed scheme that describes the pH-induced functional phase space of HSA. View Full-Text
Keywords: human serum albumin; pH; ESR/EPR; DEER; molten globule; conformational isomers; functional phase space; DLS; fluorescence human serum albumin; pH; ESR/EPR; DEER; molten globule; conformational isomers; functional phase space; DLS; fluorescence
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Reichenwallner, J.; Oehmichen, M.-T.; Schmelzer, C.E.H.; Hauenschild, T.; Kerth, A.; Hinderberger, D. Exploring the pH-Induced Functional Phase Space of Human Serum Albumin by EPR Spectroscopy. Magnetochemistry 2018, 4, 47.

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