A Natively Monomeric Deubiquitinase UCH-L1 Forms Highly Dynamic but Defined Metastable Oligomeric Folding Intermediates

Oligomerization of misfolded protein species is implicated in many human disorders. Here we showed by size-exclusion chromatography-coupled multiangle light scattering (SEC-MALS) and small-angle X-ray scattering (SEC-SAXS) that urea-induced folding intermediate of human ubiquitin C-terminal hydrolase, UCH-L1, can form well-defined dimers and tetramers under denaturing conditions despite being highly disordered. Introduction of a Parkinson disease-associated mutation, I93M, resulted in increased aggregation propensity and formation of irreversible precipitants in the presence of a moderate amount of urea. Since UCH-L1 exhibits highly populated partially unfolded forms under native conditions that resemble urea-induced folding intermediates, it is likely that these metastable dimers and tetramers can form under physiological conditions. Our findings highlighted the unique strength of integrated SEC-MALS/SAXS in quantitative analyses of the structure and dynamics of oligomeric folding intermediates that enabled us to extract information that is inaccessible to conventional biophysical techniques.