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The circulardichroism spectra (cd) of native and (partially) urea-denaturated C-phycocyanin (PC) in the spectral range 700-210 nm are presented. The large ellipticities observed in the chromophoric region of native PC are retained in the fully denatured state (8 ᴍ urea). This is similar to the behavior of the red form of phytochrome (P r ), but in contrast to C-phycoerythrin (PE). These differences are rationalized in terms of epimeric equilibria between P- and M-helix shaped chromophores. Depending on the number and location of the chirality centers present in the tetrapyrrol moities a priori, the excess populations of the inherently chiral P- and M-helices differ, thus accounting for large (PC, P r ) or small (PE) ellipticities in the denatured pigments. Hence, the large optical activity observed for the former is generated by an excess population of the P-helix induced by the asymmetric C-2, C-3 and C-3'. In PE the additional chirality center at C-16 counteracts the influence of the others. The excess population of the inherently chiral species is therefore lowered, in agreement with the nearly vanishing cd reported for denatured PE.

Circulardichroism of C-Phycocyanin: Origin of Optical Activity in Denatured Biliproteins and Evidence for an Intermediate during Unfolding