A new bright green-emitting fluorescent protein--engineered monomeric and dimeric forms

FEBS J. 2010 Apr;277(8):1967-78. doi: 10.1111/j.1742-4658.2010.07618.x. Epub 2010 Mar 18.

Abstract

Fluorescent proteins have become essential tools in molecular and biological applications. Here, we present a novel fluorescent protein isolated from warm water coral, Cyphastrea microphthalma. The protein, which we named vivid Verde fluorescent protein (VFP), matures readily at 37 degrees C and emits bright green light. Further characterizations revealed that VFP has a tendency to form dimers. By creating a homology model of VFP, based on the structure of the red fluorescent protein, DsRed, we were able to make mutations that alter the protein's oligomerization state. We present two proteins, mVFP and mVFP1, that are both exclusively monomeric, and one protein, dVFP, which is dimeric. We characterized the spectroscopic properties of VFP and its variants in comparison with enhanced green fluorescent protein (EGFP), a widely used variant of GFP. All the VFP variants are at least twice as bright as EGFP. Finally, we demonstrated the effectiveness of the VFP variants in both in vitro and in vivo detection applications.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Anthozoa / metabolism
  • Conserved Sequence
  • Dimerization
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Engineering*
  • Red Fluorescent Protein
  • Sequence Homology, Amino Acid
  • Spectrometry, Fluorescence

Substances

  • Luminescent Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins

Associated data

  • GENBANK/FN597286