Occurrence and stability of hetero-hexamer associations formed by β-carboxysome CcmK shell components

English version

Autoři: Luis F. Garcia-Alles ^aff001; Katharina Root ^aff002; Laurent Maveyraud ^aff003; Nathalie Aubry ^aff001; Eric Lesniewska ^aff004; Lionel Mourey ^aff003; Renato Zenobi ^aff002; Gilles Truan ^aff001
Působiště autorů: Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRA, INSA, Toulouse, France ^aff001; Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland ^aff002; Institut de Pharmacologie et Biologie Structurale, IPBS, Université de Toulouse, CNRS, UPS, Toulouse, France ^aff003; ICB UMR CNRS 6303, University of Bourgogne Franche-Comte, Dijon, France ^aff004
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0223877

Souhrn

The carboxysome is a bacterial micro-compartment (BMC) subtype that encapsulates enzymatic activities necessary for carbon fixation. Carboxysome shells are composed of a relatively complex cocktail of proteins, their precise number and identity being species dependent. Shell components can be classified in two structural families, the most abundant class associating as hexamers (BMC-H) that are supposed to be major players for regulating shell permeability. Up to recently, these proteins were proposed to associate as homo-oligomers. Genomic data, however, demonstrated the existence of paralogs coding for multiple shell subunits. Here, we studied cross-association compatibilities among BMC-H CcmK proteins of Synechocystis sp. PCC6803. Co-expression in Escherichia coli proved a consistent formation of hetero-hexamers combining CcmK1 and CcmK2 or, remarkably, CcmK3 and CcmK4 subunits. Unlike CcmK1/K2 hetero-hexamers, the stoichiometry of incorporation of CcmK3 in associations with CcmK4 was low. Cross-interactions implicating other combinations were weak, highlighting a structural segregation of the two groups that could relate to gene organization. Sequence analysis and structural models permitted the localization of interactions that would favor formation of CcmK3/K4 hetero-hexamers. The crystallization of these CcmK3/K4 associations conducted to the elucidation of a structure corresponding to the CcmK4 homo-hexamer. Yet, subunit exchange could not be demonstrated in vitro. Biophysical measurements showed that hetero-hexamers are thermally less stable than homo-hexamers, and impeded in forming larger assemblies. These novel findings are discussed within the context of reported data to propose a functional scenario in which minor CcmK3/K4 incorporation in shells would introduce sufficient local disorder as to allow shell remodeling necessary to adapt rapidly to environmental changes.

Klíčová slova:

Crystal structure – Signal processing – Monomers – Electrostatics – Crystallization – Stoichiometry – Operons – Macromolecular engineering

Zdroje

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Occurrence and stability of hetero-hexamer associations formed by β-carboxysome CcmK shell components

Souhrn

Klíčová slova:

Zdroje

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