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Use of Green Fluorescent Protein (GFP) to Study Cellular Dynamics

Constructing GFP-Tagged Motor Enzymes
  • Hong Cao
  • Heather Thompson
  • Eugene W. Krueger
  • Mark McNiven
Part of the Methods in Molecular Biology™ book series (MIMB, volume 161)

Abstract

The green fluorescent protein (GFP) from the Pacific Northwest jellyfish Aequorea victoria is a member of a small but important class of proteins that exhibit strong visible fluorescence without the requirement of cofactors or other enzymes. In vivo, the photoprotein aequorin binds dimers of GFP allowing the blue light emitted by aequorin to be converted to green light (1). This visible fluorescence emitted by GFP is made possible by an internal p-hydroxybenzylideneimidazolinone chromophore formed post-translationally by cyclization of Ser65, Tyr66, and Gly67 and 1,2-dehydrogenation of the tyrosine. A number of mutations have been made in GFP, most of which result in a partial or complete loss of fluorescence without significant change in relative absorption or emission peaks. These mutations probably cause misfolding of the protein, failure of chromophore formation, or quenching of the fluorescence by insufficient shielding. A few mutations, however, have resulted in enhanced GFP spectra, increased brightness, a slower rate of photobleaching, or a spectral shift resulting in the emission of a different color, such as blue, yellow, or cyan. These GFP variants emitting new colors are thus called blue fluorescent protein (BFP), yellow fluorescent protein (YFP), and cyan fluorescent protein (CFP) (2).

Keywords

Green Fluorescent Protein Polymerase Chain Reaction Primer Cyan Fluorescent Protein Polymerase Chain Reaction Machine Green Fluorescent Protein Variant 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Hong Cao
    • 1
  • Heather Thompson
    • 2
  • Eugene W. Krueger
    • 3
  • Mark McNiven
    • 3
  1. 1.Center for Basic Research in Digestive Diseases,Department of Gastroenterology and HepatologyMayo ClinicRochester
  2. 2.Department of Biochemistry and Molecular Biology,Mayo Graduate SchoolMayo Clinic and FoundationRochester
  3. 3.Center for Basic Research in Digestive Diseases,Department of Gastroenterology and HepatologyMayo Clinic and FoundationRochester

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