Peridinin–chlorophyll–protein reconstituted with chlorophyll mixtures: Preparation, bulk and single molecule spectroscopy

Brotosudarmo, T.H.P.; Hofmann, E.; Hiller, R.G.; Wörmke, S.; Mackowski, S.; Zumbusch, A.; Bräuchle, C.; Scheer, H.

doi:10.1016/j.febslet.2006.08.049

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Volume 580, Issue 22 , Pages 5257-5262, 2 October 2006

Peridinin–chlorophyll–protein reconstituted with chlorophyll mixtures: Preparation, bulk and single molecule spectroscopy

Edited by Miguel De la Rosa

T.H.P. Brotosudarmo
- Department Biologie I – Bereich Botanik, Ludwig-Maximilians-Universität München, Menzinger Str. 67, D-80638 München, Germany
- Department Chemie und Biochemie – Physikalische Chemie, and Center for Nanoscience, Ludwig-Maximilians Universität, München, Germany
E. Hofmann
- Fakultät für Biologie, Biophysik, Ruhr Universität Bochum, Germany
R.G. Hiller
- Macquarie University, Sydney, Australia
S. Wörmke
- Department Chemie und Biochemie – Physikalische Chemie, and Center for Nanoscience, Ludwig-Maximilians Universität, München, Germany
S. Mackowski
- Department Chemie und Biochemie – Physikalische Chemie, and Center for Nanoscience, Ludwig-Maximilians Universität, München, Germany
A. Zumbusch
- Fachbereich Chemie, Universität Konstanz, Germany
C. Bräuchle
- Department Chemie und Biochemie – Physikalische Chemie, and Center for Nanoscience, Ludwig-Maximilians Universität, München, Germany
H. Scheer
- Department Biologie I – Bereich Botanik, Ludwig-Maximilians-Universität München, Menzinger Str. 67, D-80638 München, Germany
- Corresponding author. Fax: +49 89 17861 271.

Received 23 July 2006; received in revised form 21 August 2006; accepted 24 August 2006. published online 04 September 2006.

Abstract

Reconstitution of the 16kDa N-terminal domain of the peridinin–chlorophyll–protein, N-PCP, with mixtures of chlorophyll a (Chl a) and Chl b, resulted in 32kDa complexes containing two pigment clusters, each bound to one N-PCP. Besides homo-chlorophyllous complexes, hetero-chlorophyllous ones were obtained that contain Chl a in one pigment cluster, and Chl b in the other. Binding of Chl b is stronger than that of the native pigment, Chl a. Energy transfer from Chl b to Chl a is efficient, but there are only weak interactions between the two pigments. Individual homo- and hetero-chlorophyllous complexes were investigated by single molecule spectroscopy using excitation into the peridinin absorption band and scanning of the Chl fluorescence, the latter show frequently well resolved emissions of the two pigments.