dc.contributor.author | Hollås, Hanne | |
dc.contributor.author | Ramirez, Juan | |
dc.contributor.author | Nominé, Yves | |
dc.contributor.author | Kostmann, Camille | |
dc.contributor.author | Toto, Angelo | |
dc.contributor.author | Gianni, Stefano | |
dc.contributor.author | Travé, Gilles | |
dc.contributor.author | Vedeler, Anni | |
dc.date.accessioned | 2023-03-13T12:35:41Z | |
dc.date.available | 2023-03-13T12:35:41Z | |
dc.date.created | 2022-12-05T14:47:44Z | |
dc.date.issued | 2022 | |
dc.identifier.issn | 0006-3495 | |
dc.identifier.uri | https://hdl.handle.net/11250/3057956 | |
dc.description.abstract | Annexins (Anxs) are a family of highly homologous proteins that bind and aggregate lipid vesicles in the presence of calcium. All members of the family contain a variable N-terminus determining specific functions, followed by a conserved core region responsible for the general calcium-dependent lipid-binding property. The core structure consists of four homologous domains (DI–DIV), each consisting of a right-handed super-helix of five α-helices. We present data from a combination of site-directed mutagenesis, NMR, and circular dichroism showing that the G25–D34 region of the N-terminus as well as the contacts between residues D38A, R63A, and Q67A of AnxA2-DI are crucial for the autonomous folding and stability of DI of AnxA2. However, we also show that the folding of the full-length protein is very robust in that mutations and truncations that disrupted the folding of AnxA2-DI did not abolish the folding of full-length AnxA2, only lowering its thermal stability. This robustness of the folding of full-length AnxA2 is likely to be mediated by the existence of at least one transient nonnative intermediate as suggested by our kinetic data using stopped-flow fluorescence experiments. We also show that hydrophobic amino acids in AnxA2-DI involved in interfacial contacts with AnxA2-DIV are important for the cooperative folding and stability of the full-length protein. Mutating all of the V57E-V98R-G101Y residues in AnxA2-DI did not affect the folding of the domain, only its stability, but prevented the cooperative folding of the full-length protein. Our collective results favor a highly cooperative and robust folding process mediated by alternative intermediate steps. Since AnxA2 is a multifunctional protein involved in several steps of the progression of cell transformation, these data on structure and folding pathways are therefore crucial to designing anticancer drugs targeting AnxA2. | en_US |
dc.language.iso | eng | en_US |
dc.publisher | Elsevier | en_US |
dc.rights | Attribution-NonCommercial-NoDerivatives 4.0 Internasjonal | * |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/deed.no | * |
dc.title | The cooperative folding of annexin A2 relies on a transient nonnative intermediate | en_US |
dc.type | Journal article | en_US |
dc.type | Peer reviewed | en_US |
dc.description.version | publishedVersion | en_US |
dc.rights.holder | Copyright 2022 Biophysical Society | en_US |
dc.source.articlenumber | 4492-4504 | en_US |
cristin.ispublished | true | |
cristin.fulltext | original | |
cristin.qualitycode | 1 | |
dc.identifier.doi | 10.1016/j.bpj.2022.10.043 | |
dc.identifier.cristin | 2088903 | |
dc.source.journal | Biophysical Journal | en_US |
dc.identifier.citation | Biophysical Journal. 2022, 121 (23), 4492-4504. | en_US |
dc.source.volume | 121 | en_US |
dc.source.issue | 3 | en_US |