Jan 7, 2011 · BamE is shown to bind specifically to phosphatidylglycerol, and comprehensive mutagenesis and interaction studies have mapped key determinants for complex binding, outer membrane integrity and cell viability, as well as revealing the role of …

β-Barrel proteins found in the outer membrane of Gram-negative bacteria serve a variety of cellular functions. Proper folding and assembly of these proteins are essential for the viability of bacteria and can also play an important role in virulence.

Sequence alignment of BamE/OmlA homologues generated by Kalign (Lassmann & Sonnhammer, 2005). C. crescentus CC1365 (Cc) was aligned with BamE/OmlA homologues that have been studied experimentally from E. coli (Ec), S. enterica serovar Typhimurium (St), P. aeruginosa (Pa) and X. axonopodis pv.

Feb 22, 2016 · In Gram-negative bacteria, the assembly of β-barrel outer-membrane proteins (OMPs) requires the β-barrel–assembly machinery (BAM) complex. We determined the crystal structure of the 200-kDa BAM...

In Escherichia coli, BAM is composed of five subunits: a β-barrel structure, BamA, and four lipoproteins, BamB–E. BamA, the core of the complex, contains five polypeptide-transport associated (POTRA) domains in the periplasm. BamA and BamD are the only essential components and are well conserved amongst Gram-negative bacteria [2, 3].

Feb 2, 2009 · BamA bears striking sequence and structural similarity to the protein-translocating TpsB proteins of the Gram-negative bacterial two-partner secretion system and to homologues in the outer ...

Jan 21, 2011 · The structure of BamE consists of a three-stranded antiparallel β-sheet packed against a pair of α-helices (Fig 1). As the ΔbamE mutant cannot grow in the presence of vancomycin, the authors identify functionally important residues of BamE by testing the effects of amino-acid substitutions in BamE on its inability to complement the growth ...

To ensure a natural folding environment, BamE was expressed in the E. coli periplasm using a PelB signal sequence (MKYLLP TAAAGLLLLAANPAMA) and with a C20S mutation to prevent amino-terminal acylation.

Outer membrane beta-barrel proteins are assembled into the membrane by the essential hetero-oligomeric BAM complex, which contains the lipoprotein BamE. We have identified a homologue of BamE, encoded by CC1365, which is located in the outer membrane of the stalked alpha-proteobacterium Caulobacter crescentus.

Mar 29, 2021 · Lipoprotein BamE was co-expressed with LolCDE, and the BamE-bound LolCDE complex was purified and reconstituted into E. coli polar lipid liposome with or without ATP and MgCl 2 (Methods)....