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Hemoglobin Fluorescence

  • Protocol
Hemoglobin Disorders

Part of the book series: Methods in Molecular Biology™ ((MIMM,volume 82))

Abstract

Protein structural analysis took a big leap forward with the discovery of aromatic amino acid and protein fluorescence (1–4). The intrinsic fluorescence of proteins is a highly sensitive reporter of conformational change at or near the fluorescent tryptophans (Trp) and tyrosines (Tyr). Phenylalanine also exhibits ultraviolet (UV) fluorescence excitation and emission, with a low quantum yield that becomes insignificant in proteins containing Tyr and Trp. The binding of specific extrinsic fluorescent probes allows the site-specific probing of other microdomains or nonfluorescent side chains. Fluorescence resonance energy transfer measurements serve as a “spectroscopic ruler” to measure intramolecular and intermolecular distances and may also be used to ascertain the magnitude of conformational change on ligand binding, protein folding, and protein-protein interactions ([5]; for basic principles of fluorescence, see ref. 6).

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Authors and Affiliations

  1. Division of Hematology, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY

    Rhoda Elison Hirstch

  2. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY

    Rhoda Elison Hirstch

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  1. Rhoda Elison Hirstch
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  1. Albert Einstein College of Medicine, Bronx, NY

    Ronald L. Nagel MD

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Hirstch, R.E. (2003). Hemoglobin Fluorescence. In: Nagel, R.L. (eds) Hemoglobin Disorders. Methods in Molecular Biology™, vol 82. Humana Press. https://doi.org/10.1385/1-59259-373-9:133

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  • DOI: https://doi.org/10.1385/1-59259-373-9:133

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-962-9

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