Title: Structural basis of BAM-mediated outer membrane β-barrel protein assembly
Chongrong Shen, Shenghai Chang, Qinghua Luo, Kevin Chun Chan, Zhibo Zhang, Bingnan Luo, Teng Xie, Guangwen Lu, Xiaofeng Zhu, Xiawei Wei, Changjiang Dong, Ruhong Zhou, Xing Zhang, Xiaodi Tang & Haohao Dong
Abstract
The outer membrane structure is common in Gram-negative bacteria, mitochondria and chloroplasts, and contains outer membrane β-barrel proteins (OMPs) that are essential interchange portals of materials. All known OMPs share the antiparallel β-strand topology4, implicating a common evolutionary origin and conserved folding mechanism. Models have been proposed for bacterial β-barrel assembly machinery (BAM) to initiate OMP folding5,6; however, mechanisms by which BAM proceeds to complete OMP assembly remain unclear. Here we report intermediate structures of BAM assembling an OMP substrate, EspP, demonstrating sequential conformational dynamics of BAM during the late stages of OMP assembly, which is further supported by molecular dynamics simulations. Mutagenic in vitro and in vivo assembly assays reveal functional residues of BamA and EspP for barrel hybridization, closure and release. Our work provides novel insights into the common mechanism of OMP assembly.