Polarized absorption spectra of green fluorescent protein single crystals: transition dipole moment directions.
Polarized absorption spectra of orthorhombic crystals of wild-type green fluorescent protein (GFP) were measured between 350 and 520 nm to obtain information on the directions of the electronic transition dipole moments ((-->)m) of the chromophore relative to the molecular axes. The transition dipole moment orientation is a basic spectroscopic parameter of relevance to biologists when interpreting Förster-type fluorescence resonance energy transfer data and for comparing absorbance and fluorescence spectra of GFP with quantum chemical calculations. Maximal extinction was obtained throughout the spectrum when the polarization direction of the electric vector of incident light was parallel to the c-axis of the crystal. The transition dipole moments were assumed to be parallel to the plane of the chromophore. With this assumption and the measured dichroic ratios in the crystals, the transition dipole moments associated with the neutral (lambda(max) = 398 nm) and anionic (lambda(max) = 478 nm) forms of the chromophore were found to subtend angles of approximately 26 degrees and 13 degrees (counterclockwise), respectively, with the vector that joins the phenolic and imidazolinone oxygen atoms of the chromophore.
Rosell
Federico I
FI
Department of Chemistry Stanford University Stanford, California 94305-5080, USA.
Boxer
Steven G
SG
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D003460:Crystallization; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins; D008027:Light; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008856:Microscopy, Fluorescence; D015591:Microspectrophotometry; D008956:Models, Chemical; D010777:Photochemistry; D011522:Protons; D037401:Scyphozoa; D012694:Serine / Q000235:genetics; D013050:Spectrometry, Fluorescence / Q000379:methods; D013912:Threonine / Q000235:genetics
D008164:Luminescent Proteins; D011522:Protons; D049452:Green Fluorescent Proteins; D013912:Threonine; D012694:Serine