title: Structure of the Haemophilus influenzae HMW1B translocator protein: evidence for a twin pore.
abstract: Secretion of the Haemophilus influenzae HMW1 adhesin occurs via the two-partner secretion pathway and requires the HMW1B outer membrane translocator. HMW1B has been subjected to extensive biochemical studies to date. However, direct examination of the structure of HMW1B has been lacking, leaving fundamental questions about the oligomeric state, the membrane-embedded beta-barrel domain, the approximate size of the beta-barrel pore, and the mechanism of translocator activity. In the current study, examination of purified HMW1B by size exclusion chromatography and negative staining electron microscopy revealed that the predominant species was a dimer. In the presence of lipid, purified HMW1B formed two-dimensional crystalline sheets. Examination of these crystals by cryo-electron microscopy allowed determination of a projection structure of HMW1B to 10 A resolution. The native HMW1B structure is a dimer of beta-barrels, with each beta-barrel measuring 40 A by 50 A in the two orthogonal directions and appearing largely occluded, leaving only a narrow pore. These observations suggest that HMW1B undergoes a large conformational change during translocation of the 125-kDa HMW1 adhesin.
authors: Biology Department, Brookhaven National Laboratory, Upton, NY 11973, USA. hli@bnl.gov
authors: Huilin
authors: H
authors: Li
authors: Susan
authors: S
authors: Grass
authors: Tao
authors: T
authors: Wang
authors: Tianbo
authors: T
authors: Liu
authors: Joseph W
authors: JW
authors: St Geme
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry* / Q000648:ultrastructure; D002850:Chromatography, Gel; D020285:Cryoelectron Microscopy; D003460:Crystallization; D019281:Dimerization; D006193:Haemophilus influenzae / Q000737:chemistry*; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D016624:Negative Staining; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary
chemical_list: D018829:Adhesins, Bacterial; C117431:HMW1 protein, bacteria; D026901:Membrane Transport Proteins
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