Quantitative interrogation of protein co-aggregation using multi-color fluorogenic protein aggregation sensors.

ABSTRACT: Co-aggregation of multiple pathogenic proteins is common in neurodegenerative diseases but deconvolution of such biochemical process is challenging. Herein, we developed a dual-color fluorogenic thermal shift assay to simultaneously report on the aggregation of two different proteins and quantitatively study their thermodynamic stability during co-aggregation. Expansion of spectral coverage was first achieved by developing multi-color fluorogenic protein aggregation sensors. Orthogonal detection was enabled by conjugating sensors of minimal fluorescence crosstalk to two different proteins via sortase-tag technology. Using this assay, we quantified shifts in melting temperatures in a heterozygous model protein system, revealing that the thermodynamic stability of wild-type proteins w

SUBMITTER: Bai Y 

PROVIDER: S-EPMC8221170 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature