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Structural characterization of the complex of SecB and metallothionein-labeled proOmpA by cryo-electron microscopy.

ABSTRACT: ProOmpA is a preprotein that is translocated across the plasma membrane by the general secretory pathway in Escherichia coli. The molecular chaperon SecB in Sec pathway can recognize and bind proOmpA for its translocation. However, the structure of the SecB/proOmpA complex remains unknown. Here, we constructed an uncleavable proOmpA fused with metallothionein at its C-terminus and labeled it with metals in vitro for the study of cryo-electron microscopy. Using single particle cryo-electron microscopy, we reconstructed 3D structure of the stable SecB/proOmpA complex. The structure shows that the major portion of preprotein locates on one side of SecB tetramer, resulting in an asymmetric binding pattern. This work also provides a possible approach to the structure determination of small protein complexes by cryo-electron microscopy.

SUBMITTER: Zhou Q

PROVIDER: S-EPMC3464278 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Structural characterization of the complex of SecB and metallothionein-labeled proOmpA by cryo-electron microscopy.

Zhou Qiang Q Sun Shan S Tai Phang P Sui Sen-Fang SF

PloS one 20121004 10

ProOmpA is a preprotein that is translocated across the plasma membrane by the general secretory pathway in Escherichia coli. The molecular chaperon SecB in Sec pathway can recognize and bind proOmpA for its translocation. However, the structure of the SecB/proOmpA complex remains unknown. Here, we constructed an uncleavable proOmpA fused with metallothionein at its C-terminus and labeled it with metals in vitro for the study of cryo-electron microscopy. Using single particle cryo-electron micro ...[more]

PMID: 23056562

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