title: Decreasing the chlorophyll a/b ratio in reconstituted LHCII: structural and functional consequences.
abstract: Trimeric (bT) and monomeric (bM) light-harvesting complex II (LHCII) with a chlorophyll a/b ratio of 0.03 were reconstituted from the apoprotein overexpressed in Escherichia coli. Chlorophyll/xanthophyll and chlorophyll/protein ratios of bT complexes and 'native' LHCII are rather similar, namely, 0.28 vs 0. 27 and 10.5 +/- 1.5 vs 12, respectively, indicating the replacement of most chlorophyll a molecules with chlorophyll b, leaving one chlorophyll a per trimeric complex. The LD spectrum of the bT complexes strongly suggests that the chlorophyll b molecules adopt orientations similar to those of the chlorophylls a that they replace. The circular dichroism (CD) spectra of bM and bT complexes indicate structural arrangements resembling those of 'native' LHCII. Thermolysin digestion patterns demonstrate that bT complexes are folded and organized like 'native' trimeric LHCII. Surprisingly, in the bT complexes at 77 K, half of the excitations that are created on either chlorophyll b or xanthophyll are transferred to chlorophyll a. No or very limited triplet transfer from chlorophyll b to xanthophyll appears to take place. However, the efficiency of triplet transfer from chlorophyll a to xanthophyll is close to 100%, even higher than in 'native' LHCII at 77 K. It is concluded from the triplet-minus-singlet and CD results that the single chlorophyll a molecule that on the average is present in each bT complex binds preferably next to a xanthophyll molecule at the interface between the monomers.
authors.affiliation: Division of Physics and Astronomy, Institute for Condensed Matter Physics and Spectroscopy, Vrije Universiteit Amsterdam, The Netherlands. foske@nat.vu.nl
authors.forename: F J
authors.initials: FJ
authors.lastname: Kleima
authors.forename: S
authors.initials: S
authors.lastname: Hobe
authors.forename: F
authors.initials: F
authors.lastname: Calkoen
authors.forename: M L
authors.initials: ML
authors.lastname: Urbanus
authors.forename: E J
authors.initials: EJ
authors.lastname: Peterman
authors.forename: R
authors.initials: R
authors.lastname: van Grondelle
authors.forename: H
authors.initials: H
authors.lastname: Paulsen
authors.forename: H
authors.initials: H
authors.lastname: van Amerongen
mesh_terms: D000042:Absorption; D002734:Chlorophyll / Q000737:chemistry* / Q000378:metabolism*; D000077194:Chlorophyll A; D002942:Circular Dichroism; D004735:Energy Transfer; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism*; D010858:Pigmentation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013329:Structure-Activity Relationship
chemical_list: D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D002734:Chlorophyll; C037184:chlorophyll b; D000077194:Chlorophyll A