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Wong, L. E.

Publications and source records attributed to Wong, L. E..

2 recordsLinked to original sources

Negatively Charged alpha-Synuclein Condensate Modulates Partitioning of Molecules

-Synuclein (Syn) aggregation via liquid-liquid phase separation has recently emerged as a crucial mechanism underlying amyloid fibril formation implicated in Parkinsons disease. However, comprehensive investigations of the physico-chemical properties of Syn condensate remains incomplete. Here, we showed that Syn condensate possesses a highly negative electrostatic potential that spans the whole condensate. This property causes differential partitioning of dye-labeled Syn as well as fluorescent molecules by an order of magnitude depending on their net charges. Consistent with this, the phase separation propensity of Syn is governed by a delicate balance between self-association of Syn and electrostatic repulsion, hence is antagonized by excess negative charge. We further demonstrated that, in differentiated neuron-like SH-SY5Y cells, Syn also forms negatively charged condensate. Our results highlighted the significant impact of Syn condensates electrostatic potential on molecular partitioning, hence calling for close examination of the electrostatic property of other biomolecular condensates.

biophysics↗

Quantitative description of the phase separation behavior of the multivalent SLP65-CIN85 complex

Biomolecular condensates play a major role in cell compartmentalization, besides membrane-enclosed organelles. The multivalent SLP65 and CIN85 proteins are downstream B cell receptor (BCR)-signaling effectors, required for a proper immune response. Both proteins phase separate together with vesicles to form pre-signaling clusters. Within this tripartite system, six PRMs of SLP65 interact promiscuously with three SH3 domains of the CIN85 monomer, establishing 18 individual SH3-PRM interactions whose individual dissociation constants we determined. Based on these 18 dissociation constants, we measured the phase separation properties of the natural SLP65/CIN85 system as well as designer constructs that emphasize the strongest SH3/PRM interactions. By modelling these various SLP65/CIN85 constructs with the program LASSI (LAttice simulation engine for Sticker and Spacer Interactions) we reproduced the observed phase separation properties. In addition, LASSI revealed a deviation in the experimental measurement, which was independently identified as a previously unknown intramolecular interaction. Thus, thermodynamic properties of the individual PRM/SH3 interactions allow to model the phase separation behavior of the SLP65/CIN85 system faithfully.

biophysics↗