Fluorescence microscopy methods in the study of protein structure and function

Heather Jensen-Smith, Benjamin Currall, Danielle Rossino, Leann Tiede, Michael Nichols, Richard Hallworth

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

As more and more proteins specific to hair cells are discovered, it becomes imperative to understand their structure and how that contributes to their function. The fluorescence microscopic methods described here can be employed to provide information on protein-protein interactions, whether homomeric or heteromeric, and on protein conformation. Here, we describe two fluorescence microscopic methodologies applied to the outer hair cell-specific membrane protein prestin: the intensity and fluorescence lifetime (FLIM) variants of FRET (Fluorescence Resonance Energy Transfer), used in the study of protein-protein interactions, and the Scanning Cysteine Accessibility Method (SCAM), used for the determination of protein conformation. The methods are readily adaptable to other proteins.

Original languageEnglish (US)
Title of host publicationAuditory and Vestibular Research
Subtitle of host publicationMethods and Protocols
PublisherHumana Press
Pages369-379
Number of pages11
ISBN (Print)9781934115626
DOIs
StatePublished - Jan 1 2009

Publication series

NameMethods in Molecular Biology
Volume493
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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  • Cite this

    Jensen-Smith, H., Currall, B., Rossino, D., Tiede, L., Nichols, M., & Hallworth, R. (2009). Fluorescence microscopy methods in the study of protein structure and function. In Auditory and Vestibular Research: Methods and Protocols (pp. 369-379). (Methods in Molecular Biology; Vol. 493). Humana Press. https://doi.org/10.1007/978-1-59745-523-7_22