Improved Isolation of SlaA and SlaB S-layer proteins in Sulfolobus acidocaldarius

Pierre Simonin (1) , Carine Lombard (1) , Arnaud Huguet (2) , Adrienne Kish (1)

1 MCAM - Molécules de Communication et Adaptation des Micro-organismes
2 METIS - Milieux Environnementaux, Transferts et Interactions dans les hydrosystèmes et les Sols

The Sulfolobus acidocaldarius S-layer is composed of two main proteins: SlaA, which forms the ordered structure of the S-layer matrix, and SlaB, which supports and anchors the S-layer into the tetraether lipid membrane. While SlaA has previously been purified by exploiting its thermotolerance and high resistance to detergents, SlaB has resisted isolation, particularly from the cell membrane. Removal of proteins other than those of the S-layer is especially difficult if large batch-scale culture volumes are unavailable. Here, we describe a benchtop-scale protocol for the purification of SlaA from S. acidocaldarius, enabling isolation of SlaB using size exclusion chromatography (gel filtration). Using this protocol, we were able to identify for the first time tetraether lipids strongly attached to SlaB via heat- and detergent-resistant interactions.

Domaines: Microbiologie et Parasitologie; Biologie moléculaire; Biochimie [q-bio.BM]

Pierre Simonin, Carine Lombard, Arnaud Huguet, Adrienne Kish. Improved Isolation of SlaA and SlaB S-layer proteins in Sulfolobus acidocaldarius. Extremophiles, 2020, 24, pp.673-680. ⟨10.1007/s00792-020-01179-9⟩. ⟨mnhn-02862458⟩