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Secondary Structure Determination of Peptides and Proteins After Immobilization

  • Protocol
Peptide Microarrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1352))

Abstract

The presentation of immobilized peptides and other small biomolecules attached to surfaces can be greatly affected by the attachment chemistry and linking moieties, resulting in altered activity and specificity. For this reason, it is critical to understand how the various aspects of surface immobilization—underlying substrate properties, tether structure, and site of linkage—affect the secondary and quaternary structures of the immobilized species. Here, we present methods for attaching cysteine-containing peptides to quartz surfaces and determining the secondary structure of surface-immobilized peptides. We specifically show that, even when covalently immobilized, changes in peptide conformation can still occur, with measurement occurring in real time.

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Notes

  1. 1.

    The RCA cleaning technique was developed during the 1960s by Werner Kern at RCA Laboratories (hence, the moniker of this solution) and has become a gold standard method for removing surface impurities from silicon semiconductors and glass substrates.

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Acknowledgments

This work was supported through the Office of Naval Research and the Naval Research Laboratory Core research programs. The views expressed herein are those of the authors and do not represent those of the US Naval Research Laboratory, the US Navy, the US Department of Defense, or the US Government.

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Authors and Affiliations

  1. US Naval Research Laboratory, Code 6900, 4555 Overlook Avenue, SW, Washington, DC, 20375, USA

    Stella H. North & Chris R. Taitt

  2. Latham & Watkins LLP, 555 Eleventh Street, NW, Washington, DC, 20004, USA

    Stella H. North

Authors
  1. Stella H. North
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  2. Chris R. Taitt
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Corresponding author

Correspondence to Chris R. Taitt .

Editor information

Editors and Affiliations

  1. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marina Cretich

  2. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marcella Chiari

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North, S.H., Taitt, C.R. (2016). Secondary Structure Determination of Peptides and Proteins After Immobilization. In: Cretich, M., Chiari, M. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 1352. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3037-1_4

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  • DOI: https://doi.org/10.1007/978-1-4939-3037-1_4

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3036-4

  • Online ISBN: 978-1-4939-3037-1

  • eBook Packages: Springer Protocols

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