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Tkachenko, O.

Publications and source records attributed to Tkachenko, O..

2 recordsLinked to original sources

αB-crystallin inhibits amyloidogenesis by disassembling aggregation nuclei

Amyloid formation is implicated in a range of neurodegenerative conditions including Alzheimers and Parkinsons diseases. The small heat-shock protein B-crystallin (BC) is associated with both, and directly inhibits amyloid formation in vitro and its toxicity in cells. Studying the mechanism of aggregation inhibition is challenging owing to sample heterogeneity and the dynamic nature of the process. Here, by means of NMR spectroscopy and chemical kinetics, we establish the mechanism by which the protein -lactalbumin aggregates and forms amyloid, and how this is inhibited by BC. In particular, we characterise the lifetime of the unstable aggregation nucleus, and determine that this species is specifically destabilised by BC. This mechanism allows the chaperone to delay the onset of aggregation, although it is overwhelmed on longer timescales. The methodology we present provides a mechanistic understanding of how BC reduces the toxicity of amyloids, and is widely applicable to other complex mixtures.

biophysics

Quantitative mass imaging of single molecules in solution

The cellular processes underpinning life are orchestrated by proteins and their interactions. Structural and dynamic heterogeneity, despite being key to protein and drug function, continues to pose a fundamental challenge to existing analytical and structural methodologies used to study these associations. Here, we use interferometric scattering microscopy to mass-image single biomolecules in solution with <2% mass error, up to 19-kDa resolution and 1-kDa precision. Thereby, we resolve oligomeric distributions at high dynamic range, detect small-molecule binding, and mass-image biomolecules composed not only of amino acids, but also heterogeneous species, such as lipo- and glycoproteins. These capabilities enable us to characterize the molecular mechanisms of processes as diverse as oligomeric selfassembly, glycoprotein cross-linking, amyloidogenic protein aggregation, and actin polymerization. Interferometric scattering mass spectrometry (iSCAMS) provides spatially resolved access to the dynamics of biomolecular interactions ranging from those involving small molecules to mesoscopic assemblies, one molecule at a time.

biophysics