Aggregation of LHCII leads to a redistribution of the triplets over the central xanthophylls in LHCII

We present laser flash-induced triplet-minus-singlet (TmSflash) and absorbance-detected-magnetic-resonance (TmSADMR) measurements on the light-harvesting chlorophyll a/b pigment-protein complex (LHCII) from pea. We investigated the influence of LHCII aggregation on xanthophyll triplet formation. The effect of aggregation was previously studied using TmSADMR [van der Vos et al. (1994) Biochim. Biophys. Acta 1208, 243-250] for LHCII from spinach, and it was concluded that aggregation leads to a large increase of the amount of intertrimer triplet transfer. However, a similar study on LHCII from pea with the use of TmSflash measurements [Barzda et al. (1998) Biochemistry 37, 546-561] showed much smaller effects. To resolve this apparent discrepancy and to compare the results of TmSADMR and TmSflash measurements, we used both techniques to study LHCII from pea, applying an identical aggregation procedure in both cases. It appears that aggregation does not lead to an increase of intertrimer triplet transfer as thought before but to a redistribution of the triplets over the two central xanthophylls (mainly lutein) that are present in each monomeric subunit of LHCII. Moreover, it is argued that the TmS band at 525 nm is due to lutein instead of violaxanthin as was reported in earlier studies. It is concluded that aggregation leads to a change in chlorophyll-xanthophyll interactions, which might explain the large change in excited-state lifetime of chlorophyll a in LHCII upon aggregation. This change in lifetime is possibly related to the phenomenon of nonphotochemical quenching in green plants, which is an important protective regulatory mechanism, that lowers the probability of photoinhibition.