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X-ray structure of a bacterial oligosaccharyltransferase

Nature volume 474, pages 350–355 (2011)Cite this article

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Abstract

Asparagine-linked glycosylation is a post-translational modification of proteins containing the conserved sequence motif Asn-X-Ser/Thr. The attachment of oligosaccharides is implicated in diverse processes such as protein folding and quality control, organism development or host–pathogen interactions. The reaction is catalysed by oligosaccharyltransferase (OST), a membrane protein complex located in the endoplasmic reticulum. The central, catalytic enzyme of OST is the STT3 subunit, which has homologues in bacteria and archaea. Here we report the X-ray structure of a bacterial OST, the PglB protein of Campylobacter lari, in complex with an acceptor peptide. The structure defines the fold of STT3 proteins and provides insight into glycosylation sequon recognition and amide nitrogen activation, both of which are prerequisites for the formation of the N-glycosidic linkage. We also identified and validated catalytically important, acidic amino acid residues. Our results provide the molecular basis for understanding the mechanism of N-linked glycosylation.

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Figure 1: Activity and structure of C. lari PglB.
Figure 2: Topology of transmembrane domain.
Figure 3: Sequon binding and recognition.
Figure 4: Catalytic site and amide nitrogen activation.
Figure 5: Proposed glycosylation mechanism of PglB.

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Protein Data Bank

Data deposits

Atomic coordinates and structure factors have been deposited with the Protein Data Bank under accession code 3RCE.

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Acknowledgements

We thank the beamline staff at the Swiss Light Source for assistance with data collection, S. Fleurkens and M. Bucher for technical assistance, and D. Arigoni for discussions. E. coli SCM6 was provided by C. Marolda and M. Valvano. C.L. was affiliated with the Life Science Zurich Graduate School. This research was supported by the NCCR Structural Biology Zurich (grant to K.P.L.) and Swiss National Science Foundation grants to M.A. (SNF 31003A-127098/1) and K.P.L. (SNF 31003A-131075/1).

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  1. Shin Numao

    Present address: Present address: Novartis Pharma AG, CH-4002 Basel, Switzerland.,

Authors and Affiliations

  1. Department of Biology, Institute of Microbiology, ETH Zurich, CH-8093 Zurich, Switzerland

    Christian Lizak, Shin Numao & Markus Aebi

  2. Department of Biology, Institute of Molecular Biology and Biophysics, ETH Zurich, CH-8093 Zurich, Switzerland

    Christian Lizak, Sabina Gerber & Kaspar P. Locher

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Contributions

C.L., M.A. and K.P.L. designed the experiments. C.L., S.G. and S.N. performed the experiments; K.P.L. performed crystallographic calculations and model building; C.L., M.A. and K.P.L. analysed the data and wrote the manuscript.

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Correspondence to Kaspar P. Locher.

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ETH Zurich has filed a patent application for the use of the PglB structure for biotechnological applications.

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Lizak, C., Gerber, S., Numao, S. et al. X-ray structure of a bacterial oligosaccharyltransferase. Nature 474, 350–355 (2011). https://doi.org/10.1038/nature10151

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