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. 1996 Oct 15;15(20):5567–5573.

Lipopolysaccharides and divalent cations are involved in the formation of an assembly-competent intermediate of outer-membrane protein PhoE of E.coli.

H de Cock 1, J Tommassen 1
PMCID: PMC452301  PMID: 8896450

Abstract

To identify the requirements for the biogenesis of outer-membrane proteins in Gram-negative bacteria, the sorting and assembly of the trimeric, pore-forming protein PhoE was studied in vitro. Purified lipopolysaccharide (LPS) in combination with low amounts of Triton X-100 and divalent cations induced the formation of folded monomers. LPS of deep-rough strains was far less efficient in the formation of folded monomers than wild-type LPS was. These folded monomers could be converted into heat-stable trimers upon addition of outer membranes and higher amounts of Triton X-100. Trimerization could precede the insertion step. These in vitro data suggest that the assembly in vivo proceeds sequentially by (i) formation of a folded monomer by interaction with LPS; (ii) sorting of the folded monomers to assembly sites in the outer membrane; (iii) trimerization; and (iv) insertion.

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