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The Best and the Brightest: Exploiting Tryptophan-Sensitized Tb3+ Luminescence to Engineer Lanthanide-Binding Tags

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Book cover Peptide Libraries

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1248))

Abstract

Consider the lanthanide metals, comprising lanthanum through lutetium. Lanthanides form stable cations with a +3 charge, and these ions exhibit a variety of useful physical properties (long-lifetime luminescence, paramagnetism, anomalous X-ray scattering) that are amenable to studies of biomolecules. The absence of lanthanide ions in living systems means that background signals are generally a nonissue; however, to exploit the advantageous properties it is necessary to engineer a robust lanthanide-binding sequence that can be appended to any macromolecules of interest. To this end, the luminescence produced by tryptophan-sensitized Tb3+ has been used as a selection marker for peptide sequences that avidly chelate these ions. A combinatorial split-and-pool library that uses two orthogonal linkers—one that is cleaved for selection and one that is cleaved for sequencing and characterization—has been used to develop lanthanide-binding tags (LBTs): peptides of 15–20 amino acids with low-nM affinity for Tb3+. Further validating the success of this screen, knowledge about LBTs has enabled the introduction of a lanthanide-binding loop in place of one of the four native calcium-binding loops within the protein calcineurin B.

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Acknowledgments

The cloning and expression of calcineurin B was done in collaboration with Prof. Patrick G. Hogan and Dr. Alina Iuga of Harvard Medical School, and their contributions are gratefully acknowledged. The authors acknowledge support for research on LBTs from the National Science Foundation (Grant MCB 0744415).

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Authors and Affiliations

  1. Department of Chemistry and Physics, Warren Wilson College, Asheville, NC, USA

    Langdon J. Martin

  2. Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA

    Barbara Imperiali

  3. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA

    Barbara Imperiali

Authors
  1. Langdon J. Martin
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  2. Barbara Imperiali
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Corresponding author

Correspondence to Barbara Imperiali .

Editor information

Editors and Affiliations

  1. Dept of Chemistry, University of Alberta, Edmonton, Alberta, Canada

    Ratmir Derda

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Martin, L.J., Imperiali, B. (2015). The Best and the Brightest: Exploiting Tryptophan-Sensitized Tb3+ Luminescence to Engineer Lanthanide-Binding Tags. In: Derda, R. (eds) Peptide Libraries. Methods in Molecular Biology, vol 1248. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2020-4_14

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  • DOI: https://doi.org/10.1007/978-1-4939-2020-4_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2019-8

  • Online ISBN: 978-1-4939-2020-4

  • eBook Packages: Springer Protocols

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