Green-fluorescent protein mutants with altered fluorescence excitation spectra

FEBS Lett. 1995 Jun 26;367(2):163-6. doi: 10.1016/0014-5793(95)00557-p.


Using random mutagenesis and visual selection of fluorescent clones, we have isolated a T203I and a E222G mutant of the Aequorea green-fluorescent protein. Each mutant has one of the two fluorescence excitation bands of the wild type deleted and retains the other without a wavelength shift. This finding is consistent with each excitation band corresponding to a distinct spectroscopic state of the chromophore. Both mutations are single amino acid exchanges which in the linear sequence are located remotely from the chromophore but in the folded protein may be situated in its vicinity. We conclude that the mutations influence the fluorescence properties by changing the interactions between the chromophore and its protein environment.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Fluorescence
  • Green Fluorescent Proteins
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / genetics
  • Molecular Sequence Data
  • Mutagenesis
  • Spectrometry, Fluorescence


  • Luminescent Proteins
  • Green Fluorescent Proteins