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Point mutations alter the mechanical stability of immunoglobulin modules

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Abstract

Immunoglobulin-like modules are common components of proteins that play mechanical roles in cells such as muscle elasticity and cell adhesion. Mutations in these proteins may affect their mechanical stability and thus may compromise their function. Using single molecule atomic force microscopy (AFM) and protein engineering, we demonstrate that point mutations in two β-strands of an immunoglobulin module in human cardiac titin alter the mechanical stability of the protein, resulting in mechanical phenotypes. Our results demonstrate a previously unrecognized class of phenotypes that may be common in cell adhesion and muscle proteins.

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Figure 1: Point mutations in an immunoglobulin module alter its mechanical stability.
Figure 2: Magnitude and kinetics of forced unfolding of mutant proteins.
Figure 3: Refolding kinetics of the I27 polyproteins.
Figure 4: Free energy diagram for the mechanical unfolding and refolding of the I27 module and its mutants.

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Acknowledgements

We thank C. Badilla-Fernandez and J. Kerkvliet for their help in polyprotein engineering. This work was supported by National Institute of Health grants to J.M.F.

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Authors and Affiliations

  1. Department of Physiology and Biophysics, Mayo Foundation, Rochester, 55905, Minnesota, USA

    Hongbin Li, Mariano Carrion-Vazquez, Andres F. Oberhauser, Piotr E. Marszalek & Julio M. Fernandez

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  1. Hongbin Li
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  2. Mariano Carrion-Vazquez
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  3. Andres F. Oberhauser
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  4. Piotr E. Marszalek
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  5. Julio M. Fernandez
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Correspondence to Julio M. Fernandez.

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Li, H., Carrion-Vazquez, M., Oberhauser, A. et al. Point mutations alter the mechanical stability of immunoglobulin modules. Nat Struct Mol Biol 7, 1117–1120 (2000). https://doi.org/10.1038/81964

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