Dynamic biomolecular condensates play crucial roles in intracellular compartmentalization and physiological functions. While engineering tools for compartmentalization have expanded add-on functionalities, directly amplifying the inherent catalytic machinery within biological phase-separated droplets has remained elusive. Herein, we developed a phase-separated oxidative folding reaction chamber based on protein disulfide isomerase A6 (PDIA6) by chemically targeting its active site CxxC motif to enhance enzymatic activity within PDIA6 droplets. A para-substituted N-methylated pyridinylmethanethiol (pMePySH) enhanced the catalytic oxidative folding of bovine pancreatic trypsin inhibitor, proinsulin, and antibody up to 12-fold within in vitro PDIA6 droplets. Furthermore, pMePySH targeted PDIA6 foci within the endoplasmic reticulum, significantly promoting insulin secretion. These findings offer a powerful platform for the spatiotemporal manipulation of protein folding, with profound implications for the scalable manufacturing of therapeutic antibodies and other complex biopharmaceuticals.
A review of Brazilian studies focusing on biomolecular condensation, discussing their main findings, experimental approaches, open questions, and interdisciplinary opportunities in the field.
A. R. Passos, A. Costa-Filho, Carolina G. Oliveira et al.· Biophysical Reviews· 0 citations
It is shown that short nucleic acids containing Gquadruplex (G4) structure can also catalyze protein folding and uncovers a previously underappreciated role for nucleic acid in proteostasis and offers a new strategy for studying nucleic acid structure-function relationship at residue level.
It is found that affinity governs the phase boundary, resistance to chemical perturbation, and molecular mobility of condensates in vitro and in human cells and is shown to be a quantitative determinant of condensate phase behavior, internal dynamics and biochemical output.
Andres Reyna, Madyson O. Briggs, Alexander F. Russell et al.· bioRxiv· 0 citations
This work aimed to generate vimentin condensates by inserting mutations mimicking posttranslational modifications associated with oxidative stress by introducing phosphomimetic residues at certain single vimentin glycosylation and/or phosphorylation sites, and suggested a modulatory role of glycosylation/phosphorylatio...
Diego Moneo-Corcuera, Paula Martínez-Cenalmor, Alma E. Martínez et al.· bioRxiv· 0 citations
Emerging high-throughput strategies to study protein condensation and aggregation at scale are reviewed, emphasizing what they truly measure, their limitations, and how the cross-talk among these complementary approaches can provide a more accurate and mechanistic mapping of sequence-to-assembly relationships.
Mariano Martín, Alice Lissmatz, Benedetta Bolognesi· Current Opinion in Structura...· 0 citations
A refolding protocol for mouse Frizzled8 cysteine-rich domain (mouse FZD8 CRD) recovered from inclusion bodies is reported, yielding a soluble, folded CRD preparation suitable for NMR analysis.
Ho-Jin Lee, Jie J. Zheng· International Journal of Mol...· 0 citations
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