Published In
Journal of Molecular Biology
Document Type
Pre-Print
Publication Date
3-2024
Subjects
Proteins -- Metabolism, Proteomics, Molecular biology, Mitosis
Abstract
Small heat shock proteins (sHSPs) are ATP-independent chaperones vital to cellular proteostasis, preventing protein aggregation events linked to various human diseases including cataract. The α-crystallins, αA-crystallin (αAc) and αB-crystallin (αBc), represent archetypal sHSPs that exhibit complex polydispersed oligomeric assemblies and rapid subunit exchange dynamics. Yet, our understanding of how this plasticity contributes to chaperone function remains poorly understood. This study investigates structural changes in αAc and αBc during client sequestration under varying degree of chaperone saturation. Using biochemical and biophysical analyses combined with single-particle electron microscopy (EM), we examined αAc and αBc in their apo-states and at various stages of client-induced co-aggregation, using lysozyme as a model client. Quantitative single-particle analysis unveiled a continuous spectrum of oligomeric states formed during the co-aggregation process, marked by significant client-triggered expansion and quasi-ordered elongation of the sHSP scaffold. These structural modifications culminated in an apparent amorphous collapse of chaperone-client complexes, resulting in the creation of co-aggregates capable of scattering visible light. Intriguingly, these co-aggregates maintain internal morphological features of highly elongated sHSP scaffolding with striking resemblance to polymeric α-crystallin species isolated from aged lens tissue. This mechanism appears consistent across both αAc and αBc, albeit with varying degrees of susceptibility to client-induced co-aggregation. Importantly, our findings suggest that client-induced co-aggregation follows a distinctive mechanistic and quasi-ordered trajectory, distinct from a purely amorphous process. These insights reshape our understanding of the physiological and pathophysiological co-aggregation processes of sHSPs, carrying potential implications for a pathway toward cataract formation.
Rights
Copyright (c) 2024 The Authors
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DOI
10.1016/j.jmb.2024.168499
Citation Details
Published as: Miller, A. P., O'Neill, S. E., Lampi, K. J., & Reichow, S. L. (2024). The α-crystallin chaperones undergo a quasi-ordered co-aggregation process in response to saturating client interaction. Journal of Molecular Biology, 168499.
Description
Pre-print author's version
This is the author’s version of a work that was accepted for publication. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published as: Miller, A. P., O'Neill, S. E., Lampi, K. J., & Reichow, S. L. (2024). The α-crystallin chaperones undergo a quasi-ordered co-aggregation process in response to saturating client interaction. Journal of Molecular Biology, 168499.