Reconstitution of the peridinin–chlorophyll a protein (PCP): evidence for functional flexibility in chlorophyll binding

Miller, David J., Catmull, Julian, Puskeiler, Robert, Tweedale, Helen, Sharples, Frank P., and Hiller, Roger G. (2005) Reconstitution of the peridinin–chlorophyll a protein (PCP): evidence for functional flexibility in chlorophyll binding. Photosynthesis Research, 86 (1-2). pp. 229-240.

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Abstract

The coding regions for the N-domain, and full length peridinin–chlorophyll a apoprotein (full length PCP), were expressed in Escherichia coli. The apoproteins formed inclusion bodies from which the peptides could be released by hot buffer. Both the above constructs were reconstituted by addition of a total pigment extract from native PCP. After purification by ion exchange chromatography, the absorbance, fluorescence excitation and CD spectra resembled those of the native PCP. Energy transfer from peridinin to Chl a was restored and a specific fluorescence activity calculated which was ~86% of that of native PCP. Size exclusion analysis and CD spectra showed that the N-domain PCP dimerized on reconstitution. Chl a could be replaced by Chl b, 3-acetyl Chl a, Chl d and Bchl using the N-domain apo protein. The specific fluorescence activity was the same for constructs with Chl a, 3-acetyl Chl a, and Chl d but significantly reduced for those made with Chl b. Reconstitutions with mixtures of chlorophylls were also made with eg Chl b and Chl d and energy transfer from the higher energy Qy band to the lower was demonstrated.

Item ID: 7020
Item Type: Article (Research - C1)
ISSN: 1573-5079
Keywords: chlorophyll; energy transfer; peridinin; protein reconstitution
Date Deposited: 01 Apr 2010 04:10
FoR Codes: 06 BIOLOGICAL SCIENCES > 0601 Biochemistry and Cell Biology > 060199 Biochemistry and Cell Biology not elsewhere classified @ 100%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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