NMR Solution Structure of the Integral Membrane Enzyme DsbB: Functional Insights into DsbB-Catalyzed Disulfide Bond Formation

Yunpeng Zhou, Tomasz Cierpicki, Ricardo H.Flores Jimenez, Stephen M. Lukasik, Jeffrey F. Ellena, David S. Cafiso, Hiroshi Kadokura, Jon Beckwith, John H. Bushweller

Research output: Contribution to journalArticlepeer-review

152 Citations (Scopus)

Abstract

We describe the NMR structure of DsbB, a polytopic helical membrane protein. DsbB, a bacterial cytoplasmic membrane protein, plays a key role in disulfide bond formation. It reoxidizes DsbA, the periplasmic protein disulfide oxidant, using the oxidizing power of membrane-embedded quinones. We determined the structure of an interloop disulfide bond form of DsbB, an intermediate in catalysis. Analysis of the structure and interactions with substrates DsbA and quinone reveals functionally relevant changes induced by these substrates. Analysis of the structure, dynamics measurements, and NMR chemical shifts around the interloop disulfide bond suggest how electron movement from DsbA to quinone through DsbB is regulated and facilitated. Our results demonstrate the extraordinary utility of NMR for functional characterization of polytopic integral membrane proteins and provide insights into the mechanism of DsbB catalysis.

Original languageEnglish
Pages (from-to)896-908
Number of pages13
JournalMolecular Cell
Volume31
Issue number6
DOIs
Publication statusPublished - 2008 Sept 26

Keywords

  • PROTEINS

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