The relationship between the binding of dicyclohexylcarbodiimide and quenching of chlorophyll fluorescence in the light-harvesting proteins of photosystem II.
The relationship between the binding of dicyclohexylcarbodiimide (DCCD) to isolated light-harvesting proteins of photosystem II and the inhibition of chlorophyll fluorescence quenching by DCCD have been investigated. For a range of different complexes an approximately linear relationship was obtained between the efficiency of DCCD binding and the DCCD-dependent reversal of fluorescence quenching. The most efficient labeling was found for the minor light-harvesting complexes, CP29 and CP26. In the case of the former, five different preparations were compared including two reconstituted complexes in which a putative DCCD-binding site had been mutagenized. Again, an approximately linear relationship between DCCD binding and the extent of reversal of fluorescence quenching was found. However, the binding of DCCD was found to occur at least an order of magnitude faster than the change in fluorescence. The results are discussed in terms of the multiplicity of DCCD-binding sites and the influence of protein structure on both the binding of DCCD and the fluorescence quenching mechanism.
Robert Hill Institute for Photosynthesis Research, Department of Molecular Biology & Biotechnology, University of Sheffield, UK.
A V
AV
Ruban
P
P
Pesaresi
U
U
Wacker
K D
KD
Irrgang
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R
Bassi
P
P
Horton
D001665:Binding Sites; D002734:Chlorophyll / Q000378:metabolism*; D004024:Dicyclohexylcarbodiimide / Q000378:metabolism*; D005454:Fluorescence Polarization; D005973:Glutamine / Q000235:genetics; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes; D016297:Mutagenesis, Site-Directed; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D013050:Spectrometry, Fluorescence; D018724:Spinacia oleracea; D014633:Valine / Q000235:genetics
C069660:CP29 light harvesting complex; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D005973:Glutamine; C070970:photosystem II, chlorophyll-binding protein, CP-47; D002734:Chlorophyll; D004024:Dicyclohexylcarbodiimide; D014633:Valine