RT Journal Article
T1 Probing supramolecular protein assembly using covalently attached fluorescent molecular rotors
A1 Kubánková, Markéta
A1 López Duarte, Ismael
A1 Bull, James A.
A1 Vadukul, Devkee M.
A1 Serpell, Louise C.
A1 Victor, Marie de Saint
A1 Stride, Eleanor
A1 Kuimova, Marina K.
AB Changes in microscopic viscosity and macromolecular crowding accompany the transition of proteinsfrom their monomeric forms into highly organised fibrillar states. Previously, we have demonstrated thatviscosity sensitive fluorophores termed ‘molecular rotors’, when freely mixed with monomers of interest,are able to report on changes in microrheology accompanying amyloid formation, and measured anincrease in rigidity of approximately three orders of magnitude during aggregation of lysozyme andinsulin. Here we extend this strategy by covalently attaching molecular rotors to several proteins capableof assembly into fibrils, namely lysozyme, fibrinogen and amyloid-b peptide (Ab(1e42)). We demonstrate that upon covalent attachment the molecular rotors can successfully probe supramolecular assembly in vitro. Importantly, our new strategy has wider applications in cellulo and in vivo, sincecovalently attached molecular rotors can be successfully delivered in situ and will colocalise with theaggregating protein, for example inside live cells. This important advantage allowed us to follow themicroscopic viscosity changes accompanying blood clotting and during Ab(1e42) aggregation in live SHSY5Y cells. Our results demonstrate that covalently attached molecular rotors are a widely applicable toolto study supramolecular protein assembly and can reveal microrheological features of aggregatingprotein systems both in vitro and in cellulo not observable through classical fluorescent probes operatingin light switch mode.
PB Elsevier
SN 0142-9612
YR 2017
FD 2017-09
LK https://hdl.handle.net/20.500.14352/114924
UL https://hdl.handle.net/20.500.14352/114924
LA eng
NO Kubánková M, López-Duarte I, Bull JA, Vadukul DM, Serpell LC, De Saint Victor M, et al. Probing supramolecular protein assembly using covalently attached fluorescent molecular rotors. Biomaterials [Internet]. septiembre de 2017 [citado 17 de enero de 2025];139:195-201. Disponible en: https://linkinghub.elsevier.com/retrieve/pii/S0142961217304027
NO Engineering & Physical Sciences Research Council
DS Docta Complutense
RD 18 abr 2025