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Synthesis of a fluorescent probe for sensing multiple protein states

Author:
Kucherak, Oleksandr A   Shvadchak, Volodymyr V   Kyriukha, Yevhenii A   Yushchenko, Dmytro A   


Journal:
European Journal of Organic Chemistry


Issue Date:
2018


Abstract(summary):

We synthesized solvatochromic thiol-reactive fluorescent probes 6FM-M and 7AFM-M possessing two-band emission. 7AFM-M was designed to have a smaller dipole moment than 6FM-M and as a result it showed better separation of the two emission bands. Both probes were used to label a cysteine mutant of -synuclein (Syn), a presynaptic neuronal protein associated with Parkinson's disease. We investigated the ability of the probes to sense Syn binding to lipid membranes and Syn fibrillization, and compared their sensitivity with that of one-band solvatochromic probes. We found that while all the tested probes can discriminate two Syn states, only the 7AFM-M probe is able to clearly discriminate all three protein states of Syn: unstructured, membrane-bound and fibrillar Syn. Finally, using Syn labelled with 7AFM-M, we demonstrated that high density of Syn on lipid membranes may lead to the partial membrane destruction with a formation of lipoprotein particles.


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