Tracking Cell Wall-anchored Proteins in Gram-positive Bacteria

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Book Chapter

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Immunofluorescence Microscopy, Cell Wall-anchored Proteins, Staphylococcus Aureus, Protein A (Spa), Surface Display, Cross-wall, Septum

Digital Object Identifier (DOI)


Cell wall-anchored surface proteins are integral components of Gram-positive bacterial cell envelope and vital for bacterial survival in different environmental niches. To fulfill their functions, surface protein precursors translocate from cytoplasm to bacterial cell surface in three sequential steps: secretion across the cytoplasmic membrane, covalently anchoring to the cell wall precursor lipid II by sortase A, and incorporation of the lipid II-linked precursors into mature cell wall peptidoglycan. Here, we describe a series of immunofluorescence microscopy methods to track the subcellular localization of cell wall-anchored proteins along the sorting pathway. While the protocols are tailored to Staphylococcus aureus, they can be readily adapted to localize cell wall-anchored proteins as well as membrane proteins in other Gram-positive bacteria.

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Citation / Publisher Attribution

Tracking Cell Wall-anchored Proteins in Gram-positive Bacteria, in Hung Ton-That (Ed.), The Bacterial Cell Wall, v. 2727, Humana, p. 193-204