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bioRxiv · 10.1101/2025.05.09.653011

A Disease-Associated Mutation Impedes PPIA Through Allosteric Dynamics Modulation

Hattori, Y.·Kumashiro, M.·Kumeta, H.·Kyo, T.·Kawagoe, S.·Matsusaki, M.·Saio, T.
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Abstract

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by motor neuron degeneration. Peptidylprolyl cis-trans isomerase A (PPIA) is a molecular chaperone involved in protein folding, and its dysfunction has been linked to ALS pathogenesis as proline is recognized as a key residue for maintaining proper folding of ALS-related proteins. A recent study identified a K76E mutation in PPIA in sporadic ALS patients, but its effects on protein function and structure remain unclear. In this study, we used biochemical and biophysical techniques to investigate the structural and functional consequences of the K76E mutation. Our results show that K76E significantly reduces enzyme activity without affecting structure, monodispersity, or substrate recognition. Significant effects of K76E mutation were identified by relaxation dispersion NMR experiments, showing that K76E disrupts key protein dynamics and alters an allosteric network essential for isomerase activity. Corroborated by theoretical kinetic analysis, this dynamics data, revealing the exchange process for K76E approximately one-tenth that of the wild type, explain the reduced cis-trans isomerase activity of the K76E mutant. These findings suggest that the pathogenic effect of K76E arises primarily from impaired protein dynamics rather than direct structural disruption. Our study provides new insights into the molecular mechanisms underlying ALS-associated mutations and their impact on protein function. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/653011v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@82f81corg.highwire.dtl.DTLVardef@6f0a04org.highwire.dtl.DTLVardef@1fabe12org.highwire.dtl.DTLVardef@d49d11_HPS_FORMAT_FIGEXP M_FIG For Table of Contents C_FIG

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Hattori, Y., Kumashiro, M., Kumeta, H., Kyo, T., Kawagoe, S., Matsusaki, M., Saio, T.. 2025-05-14. A Disease-Associated Mutation Impedes PPIA Through Allosteric Dynamics Modulation. https://doi.org/10.1101/2025.05.09.653011

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