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Dynamic local unfolding in the serpin α-1 antitrypsin provides a mechanism for loop insertion and polymerization

Nature Structural & Molecular Biology volume 18, pages 222–226 (2011)Cite this article

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Abstract

The conformational plasticity of serine protease inhibitors (serpins) underlies both their activities as protease inhibitors and their susceptibility to pathogenic misfolding and aggregation. Here, we structurally characterize a sheet-opened state of the serpin α-1 antitrypsin (α1AT) and show how local unfolding allows functionally essential strand insertion. Mutations in α1AT that cause polymerization-induced serpinopathies map to the labile region, suggesting that the evolution of serpin function required sampling of high risk conformations on a dynamic energy landscape.

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Figure 1: Design of cysteine mutations to probe low denaturant–induced strand opening in the serpin α1AT.
Figure 2: Accessibility of cysteinyl thiols at the indicated position in α1AT single-cysteine variants as a function of denaturant and comparison to unfolding monitored by CD.
Figure 3: Structural context of cysteines belonging to the three classes with varying extents of PEGylation.
Figure 4: Solvent accessibility of single cysteines at the indicated positions in the Z variant of α1AT.
Figure 5: Local unfolding leads to the sheet-opened intermediate of α1AT.

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Acknowledgements

We thank C.L. Cooney (Massachusetts Institute of Technology) for providing a modified form of plasmid pEAT8 (see Online Methods), originally from M.H. Yu (Korea Institute of Science and Technology). We acknowledge S. Eyles and the University of Massachusetts, Amherst, mass spectrometry facility for ESI-MS results, and we thank E. Clerico, D. Hebert and A. Gershenson for stimulating discussions and critical reading of the manuscript. This work was supported by grants from the US National Institutes of Health (OD-00045 to L.M.G.) and the Alpha-1 Foundation (to B.K.).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, Massachusetts, USA

    Beena Krishnan & Lila M Gierasch

  2. Department of Chemistry, University of Massachusetts Amherst, Amherst, Massachusetts, USA

    Lila M Gierasch

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  1. Beena Krishnan
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Contributions

B.K. and L.M.G. designed the experiments, B.K. carried out the experiments, B.K. and L.M.G. interpreted results and B.K. and L.M.G. wrote the manuscript.

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Correspondence to Lila M Gierasch.

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Krishnan, B., Gierasch, L. Dynamic local unfolding in the serpin α-1 antitrypsin provides a mechanism for loop insertion and polymerization. Nat Struct Mol Biol 18, 222–226 (2011). https://doi.org/10.1038/nsmb.1976

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