crystal structure of bamb from pseudomonas aeruginosa and functional evaluation of its conserved structural features晶体结构的bamb铜绿假单胞菌的守恒的结构特点和功能评价.pdfVIP

Crystal Structure of BamB from Pseudomonas aeruginosa and Functional Evaluation of Its Conserved Structural Features ˇ Katarina Bartos Jansen, Susan Lynn Baker, Marcelo Carlos Sousa* Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, Colorado, United States of America Abstract The assembly of b-barrel Outer Membrane Proteins (OMPs) in the outer membrane is essential for Gram-negative bacteria. The process requires the b-Barrel Assembly Machine (BAM), a multiprotein complex that, in E. coli, is composed of the OMP BamA and four lipoproteins BamB-E. Whereas BamA and BamD are essential, deletion of BamB, C or E produce membrane permeability defects. Here we present the high-resolution structure of BamB from Pseudomonas aeruginosa. This protein can complement the deletion of bamB in E. coli indicating that they are functionally equivalent. Conserved structural features ˚ include an eight-bladed b-propeller fold stabilized by tryptophan docking motifs with a central pore about 8 A in diameter at the narrowest point. This pore distinguishes BamB from related b-propellers, such as quinoprotein dehydrogenases. However, a double mutation designed to block this pore was fully functional indicating that the opening is not essential. Two loops protruding from the bottom of the propeller are conserved and mediate binding to BamA. Conversely, an additional loop only present in E. coli BamB is not required for function. A cluster of highly conserved residues in a groove between blades 6 and 7 is crucial for proper BamB folding or biogenesis. It has been proposed that BamB may bind nascent OMPs by b-augmentation to its propeller outer strands, or recognize the aromatic resi