Fluorescence polarization studies on four biliproteins and a bilin model for phycoerythrin 545.
Fluorescence (excitation) polarization spectroscopy in the wavelength region of the bilin chromophores was applied to phycoerythrocyanin (CV-phycocyanin), phycocyanins 645 and 612, and phycoerythrin 545. The cryptomonad biliproteins - phycoerythrin 545 and phycocyanins 612 and 645 - were studied as both protein dimers having an alpha(2)beta(2) polypeptide structure and as alphabeta monomers. The cyanobacterial phycoerythrocyanin (CV-phycocyanin) was a trimeric oligomer. The changes in polarization across the spectrum were attributed to transfers of energy between bilins. Cryptomonad biliproteins are isolated as dimers. The similarities between their steady-state fluorescence polarization spectra and those of the corresponding monomers suggested that the monomers' conformations were analogous to the dimers. This supports the use of monomers in the study of dimer bilin organization. The unusual polarization spectrum of phycoerythrin 545 was explained using a model for the topography of its bilins. Obtaining the emission spectra of phycoerythrin 545 at several temperatures and a deconvolution of the dimer circular dichroism spectrum also successfully tested the bilin model. Circular dichroism spectroscopy was used to determine which polarization changes are formed by Förster resonance energy transfers and which may be produced by internal conversions between high- and low-energy states of pairs of exciton-coupled bilins. Attempts were made to assign energy transfer events to the corresponding changes in fluorescence polarization for each of the four biliproteins.
MacColl
R
R
Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY, USA. robert.maccoll@wadsworth.org
Eisele
L E
LE
Marrone
J
J
D001654:Bile Pigments / Q000737:chemistry*; D002942:Circular Dichroism; D019281:Dimerization; D005454:Fluorescence Polarization; D052980:Phycobilins; D010798:Phycocyanin / Q000031:analogs & derivatives / Q000737:chemistry; D010799:Phycoerythrin / Q000737:chemistry*
D001654:Bile Pigments; D052980:Phycobilins; C053456:phycocyanin 612; C039637:phycocyanin 645; C045714:phycoerythrin 545; C038397:phycoerythrocyanin; D010798:Phycocyanin; D010799:Phycoerythrin