Probing the pigment binding sites in LHCII with resonance Raman spectroscopy: The effect of mutations at S123. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue BBA - Biochimica et Biophysica Acta Année : 2016

Probing the pigment binding sites in LHCII with resonance Raman spectroscopy: The effect of mutations at S123.

Résumé

Resonance Raman spectroscopy was used to evaluate the structure of light-harvesting chlorophyll (Chl) a/b complexes of photosystem II (LHCII), reconstituted from wild-type (WT) and mutant apoproteins over-expressed in Escherichia coli. The point mutations involved residue S123, exchanged for either P (S123P) or G (S123G). In all reconstituted proteins, lutein 2 displayed a distorted conformation, as it does in purified LHCII trimers. Reconstituted WT and S123G also exhibited a conformation of bound neoxanthin (Nx) molecules identical to the native protein, while the S123P mutation was found to induce a change in Nx conformation. This structural change of neoxanthin is accompanied by a blue shift of the absorption of this carotenoid molecule. The interactions assumed by (and thus the structure of the binding sites of) the bound Chls b were found identical in all the reconstituted proteins, and only marginally perturbed as compared to purified LHCII. The interactions assumed by bound Chls a were also identical in purified LHCII and the reconstituted WT. However, the keto carbonyl group of one Chl a, originally free-from-interactions in WT LHCII, becomes involved in a strong H-bond with its environment in LHCII reconstituted from the S123P apoprotein. As the absorption in the Qy region of this protein is identical to that of the LHCII reconstituted from the WT apoprotein, we conclude that the interaction state of the keto carbonyl of Chl a does not play a significant role in tuning the binding site energy of these molecules.

Dates et versions

hal-01356088 , version 1 (24-08-2016)

Identifiants

Citer

Elizabeth Kish, Ke Wang, Manuel J Llansola-Portoles, Cristian Ilioaia, Andrew A Pascal, et al.. Probing the pigment binding sites in LHCII with resonance Raman spectroscopy: The effect of mutations at S123.. BBA - Biochimica et Biophysica Acta, 2016, 1857 (9), pp.1490-6. ⟨10.1016/j.bbabio.2016.06.001⟩. ⟨hal-01356088⟩
63 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More