VELOURS, Christophe; ZHOU, Jingji; ZECCHIN, Paolo; HE, Nisha; SALAMEH, Myriam; GOLINELLI, Marie-Pierre; GOLINELLI-PIMPANEAU, Béatrice

doi:10.3390/biom12020270

VELOURS, Christophe
Microbiologie Fondamentale et Pathogénicité [MFP]

ZHOU, Jingji

ZECCHIN, Paolo

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Ce document a été publié dans

Biomolecules. 2022-02-09

Résumé en anglais

Size Exclusion Chromatography coupled with Multi-Angle Light Scattering (SEC-MALS) is a technique that determines the absolute molar mass (molecular weight) of macromolecules in solution, such as proteins or polymers, by detecting their light scattering intensity. Because SEC-MALS does not rely on the assumption of the globular state of the analyte and the calibration of standards, the molar mass can be obtained for proteins of any shape, as well as for intrinsically disordered proteins and aggregates. Yet, corrections need to be made for samples that absorb light atthe wavelength of the MALS laser, such as iron–sulfur [Fe-S] cluster-containing proteins. We analyze several examples of [2Fe-2S] and [4Fe-4S] cluster-containing proteins, for which various corrections were applied to determine the absolute molar mass of both the apo- and holo-forms. Importantly, the determination of the absolute molar mass of the [2Fe-2S]-containing holo-NEET proteins allowed us to ascertain a change in the oligomerization state upon cluster binding and, thus, to highlight one essential function of the cluster.< Réduire

Mots clés en anglais

SEC-MALS

size exclusion chromatography

multi-angle light scattering

molar mass

molecular weight

Fe-S cluster

iron-sulfur cluster

[Fe-S]-binding protein

NEET protein

URI

https://oskar-bordeaux.fr/handle/20.500.12278/182125

DOI

http://dx.doi.org/10.3390/biom12020270

Unités de recherche

MFP (Laboratoire Microbiologie Fondamentale et Pathogénicité) - UMR 5234

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Determination of the Absolute Molar Mass of [Fe-S]-Containing Proteins Using Size Exclusion Chromatography-Multi-Angle Light Scattering (SEC-MALS)

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Résumé en anglais

Mots clés en anglais

URI

DOI

Unités de recherche