Exciton splitting in phycoerythrin 545.

Abstract

Phycoerythrin 545 is a biliprotein having a polypeptide structure of alpha 2 beta 2, and each alpha and beta polypeptide has chromophores. Circular dichroism (CD) and absorption spectroscopy in the visible region together with various biochemical protocols have been used to study these chromophores. The CD spectrum exhibits overlapping positive and negative bands. Exciton splitting between closely-spaced pairs of chromophores produces a CD spectrum that has positive and negative bands of equal rotational strengths, a conservative spectrum. Alternatively, any positive or negative band could arise from a single chromophore. The results of this study demonstrate that exciton splitting is the likely cause of the negative and corresponding positive bands. The CD spectra of the separated alpha and beta polypeptides, under conditions where the polypeptide structure is denatured, have no negative bands. When the polypeptides are allowed to refold individually, the chromophores on the beta polypeptide regain a combination of negative and positive CD bands. The spectrum of the alpha polypeptide shows no evidence of exciton splitting under these refolding conditions. In another approach, urea is added to the protein in low concentrations, which result in changes in the conformation and perhaps association of the protein. A difference CD spectrum of native protein minus protein in 0.8 M urea shows a spectrum characteristic of exciton splitting. Moreover, the remaining CD spectra in 0.8, 1.6, or 2.4 M urea still show the possibility of further exciton splitting, but a slightly different wavelengths from the spectrum that is deleted by 0.8 M urea. This finding may suggest that there are two types of exciton splitting.