Pyron, R. | bioRxiv Science

bioRxiv2026

Anchored Brownian motion and Bayesian methods for the analysis of single particle tracking data

We present a novel statistical method and a prototype computational implementation for estimating the diffusion coefficient from single particle tracking (SPT) data. Our method is based on anchored Brownian motion which is a novel representation that relaxes the restrictions of conventional Brownian motion. Our method is fully developed in Bayesian terms and allows for robust estimation of diffusion coefficient and quantification of the uncertainly propagated from limited data quantity and quality as appropriate for the analysis of live-cell SPT data. We compare our methods with conventional Brownian motion and demonstrate superior performance in estimating the correct value of the diffusion coefficient. Finally, we benchmark our methods with SPT data from in cellulo and in silico experiments.

biophysics↗

bioRxiv2024

Cholesterol inhibits oncogenic EphA2 activation

The receptor tyrosine kinase EphA2 drives cancer malignancy by facilitating metastasis. EphA2 can be found in different self-assembly states: as a monomer, dimer, and oligomer. However, our understanding remains limited regarding which EphA2 state is responsible for driving pro-metastatic signaling. To address this limitation, we have developed SiMPull-POP, a single-molecule method for accurate quantification of membrane protein self-assembly. Our experiments revealed that a reduction of plasma membrane cholesterol strongly promoted EphA2 self-assembly. Indeed, low cholesterol caused a similar effect to the EphA2 ligand ephrinA1-Fc. These results indicate that cholesterol inhibits EphA2 assembly. Phosphorylation studies in different cell lines revealed that low cholesterol increased phospho-serine levels, the signature of oncogenic signaling. Investigation of the mechanism that cholesterol uses to inhibit the assembly and activity of EphA2 indicate an in-trans effect, where EphA2 is phosphorylated by protein kinase A downstream of beta-adrenergic receptor activity, which cholesterol also inhibits. Our study not only provides new mechanistic insights on EphA2 oncogenic function, but also suggests that cholesterol acts as a molecular safeguard mechanism that prevents uncontrolled self-assembly and activation of EphA2.

biochemistry↗