Issue 1, 2025

An albumin unfolding and refolding cycle induced by a time-controlled pH jump

Abstract

Given the intimate connection between the structure and function of biological macromolecules, the ability to temporally control their unfolding–refolding process enables temporal regulation over specific functionalities, potentially applicable in innovative domains, including the construction of protein-based actuators or programmable catalysis and drug release in complex biotechnological processes. We show here how a temporally controlled protein unfolding–refolding cycle can be coupled in time with programmed pH sequences achieved through the spontaneous decomposition of an activated carboxylic acid. Specifically, we illustrate this process at the molecular level using albumin, the most prevalent protein found in plasma, for which a temporary shift from native to unfolded forms is promoted using nitroacetic acid, able to undergo base-catalysed decarboxylation when solubilized in water solution. As detected by small angle X-ray scattering and intrinsic tryptophan fluorescence, starting from the protein in its native form, the acid addition triggers unfolding to a partially denatured state and a subsequent time-tunable pH rise with gradual refolding that recapitulates the intermediate steps detected at the same pH values by static acidification, fitting within a framework of full reversibility of the structural changes as a function of the protein protonation state. At the end of the pH jump, the native structure is fully recovered, making this method a chemical tool to achieve a complete protein conformational sequence programmed in the timeframe of minutes without further intervention.

Graphical abstract: An albumin unfolding and refolding cycle induced by a time-controlled pH jump

Supplementary files

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Article information

Article type
Paper
Submitted
03 Aug 2024
Accepted
25 Sep 2024
First published
26 Sep 2024
This article is Open Access
Creative Commons BY license

Org. Biomol. Chem., 2025,23, 118-125

An albumin unfolding and refolding cycle induced by a time-controlled pH jump

A. Del Giudice, D. Del Giudice, E. Spatola, V. Alemanno, L. Galantini and S. Di Stefano, Org. Biomol. Chem., 2025, 23, 118 DOI: 10.1039/D4OB01289E

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