Configuration and dynamics of xanthophylls in light-harvesting antennae ofhigher plants - Spectroscopic analysis of isolated light-harvesting complex of photosystem II and thylakoid membranes

Resonance Raman excitation spectroscopy combined with ultra low temperature absorption spectral analysis of the major xanthophylls of higher plants in isolated antenna and intact thylakoid membranes was used to identify carot enoid absorption regions and study their molecular configuration. The major electronic transitions of the light-harvesting complex of photosystem II ( LHCIIb) xanthophylls have been identified for both the monomeric and trimer ic states of the complex. One long wavelength state of lutein with a 0-0 tr ansition at 510 nm was detected in LHCIIb trimers, The short wavelength 0-0 transitions of lutein and neoxanthin were located at 495 and 486 nm, respe ctively. In monomeric LHCIIb, both luteins absorb around 495 nm, but slight differences in their protein environments give rise to a broadening of thi s band. The resonance Raman spectra of violaxanthin and zeaxanthin in intac t thylakoid membranes was determined. The broad 0-0 absorption transition f or zeaxanthin was found to be located in the 503-511 nm region. Violaxanthi n exhibited heterogeneity, having two populations with one absorbing at 497 nm (0-0), 460 nm (0-1), and 429 nm (0-2), and the other major pool absorbi ng at 488 nm (0-0), 452 nm (0-1), and 423 nm (0-2), The origin of this hete rogeneity is discussed. The configuration of zeaxanthin and violaxanthin in thylakoid membranes was different from that of free pigments, and both xan thophylls (notably, zeaxanthin) were found to be well coordinated within th e antenna proteins in vivo, arguing against the possibility of their free d iffusion in the membrane and supporting our recent biochemical evidence of their association with intact oligomeric light-harvesting complexes.