Divide, conquer and reconstruct: How to solve the 3D structure of recalcitrant Micro-Exon Gene (MEG) protein from Schistosoma mansoni.

Autor: Nedvedova S; Université de Lyon, CNRS, UCB Lyon1, Institut des Sciences Analytiques, UMR5280, 5 rue de la Doua, Villeurbanne, France.; Department of Chemistry, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague, Czech Republic.; Department of Zoology and Fisheries, Center of Infectious Animal Diseases, Czech University of Life Sciences Prague, Prague, Czech Republic., Guillière F; Université de Lyon, CNRS, UCB Lyon1, Institut des Sciences Analytiques, UMR5280, 5 rue de la Doua, Villeurbanne, France., Miele AE; Université de Lyon, CNRS, UCB Lyon1, Institut des Sciences Analytiques, UMR5280, 5 rue de la Doua, Villeurbanne, France.; Department of Biochemical Sciences, Sapienza University of Rome, Rome, Italy., Cantrelle FX; Université de Lille, CNRS, UMR8576 -UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, Lille, France., Dvorak J; Department of Zoology and Fisheries, Center of Infectious Animal Diseases, Czech University of Life Sciences Prague, Prague, Czech Republic.; Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Prague, Czechia.; Faculty of Environmental Sciences, Center of Infectious Animal Diseases, Czech University of Life Sciences in Prague, Prague, Czech Republic., Walker O; Université de Lyon, CNRS, UCB Lyon1, Institut des Sciences Analytiques, UMR5280, 5 rue de la Doua, Villeurbanne, France., Hologne M; Université de Lyon, CNRS, UCB Lyon1, Institut des Sciences Analytiques, UMR5280, 5 rue de la Doua, Villeurbanne, France.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2023 Aug 03; Vol. 18 (8), pp. e0289444. Date of Electronic Publication: 2023 Aug 03 (Print Publication: 2023).
DOI: 10.1371/journal.pone.0289444
Abstrakt: Micro-Exon Genes are a widespread class of genes known for their high variability, widespread in the genome of parasitic trematodes such as Schistosoma mansoni. In this study, we present a strategy that allowed us to solve the structures of three alternatively spliced isoforms from the Schistoma mansoni MEG 2.1 family for the first time. All isoforms are hydrophobic, intrinsically disordered, and recalcitrant to be expressed in high yield in heterologous hosts. We resorted to the chemical synthesis of shorter pieces, before reconstructing the entire sequence. Here, we show that isoform 1 partially folds in a-helix in the presence of trifluoroethanol while isoform 2 features two rigid elbows, that maintain the peptide as disordered, preventing any structuring. Finally, isoform 3 is dominated by the signal peptide, which folds into a-helix. We demonstrated that combining biophysical techniques, like circular dichroism and nuclear magnetic resonance at natural abundance, with in silico molecular dynamics simulation for isoform 1 only, was the key to solve the structure of MEG 2.1. Our results provide a crucial piece to the puzzle of this elusive and highly variable class of proteins.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2023 Nedvedova et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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