Construction and characterization of genetically modified Synechocystis sp. PCC 6803 photosystem II core complexes containing carotenoids with shorter π-conjugation than β-carotene

James A. Bautista, Cara A. Tracewell, Eberhard Schlodder, Francis X. Cunningham, Gary W Brudvig, Bruce A. Diner

Research output: Contribution to journalArticle

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Abstract

β-Carotene has been identified as an intermediate in a secondary electron transfer pathway that oxidizes Chlz and cytochrome b 559 in Photosystem II (PS II) when normal tyrosine oxidation is blocked. To test the redox function of carotenoids in this pathway, we replaced the ζ-carotene desaturase gene (zds) or both the zds and phytoene desaturase (pds) genes of Synechocystis sp. PCC 6803 with the phytoene desaturase gene (crtI) of Rhodobacter capsulatus, producing carotenoids with shorter conjugated π-electron systems and higher reduction potentials than β-carotene. The PS II core complexes of both mutant strains contain approximately the same number of chlorophylls and carotenoids as the wild type but have replaced β-carotene (11 double bonds), with neurosporene (9 conjugated double bonds) and β-zeacarotene (9 conjugated double bonds and 1 β-ionylidene ring). The presence of the ring appears necessary for PS II assembly. Visible and near-infrared spectroscopy were used to examine the light-induced formation of chlorophyll and carotenoid radical cations in the mutant PS II core complexes at temperatures from 20 to 160 K. At 20 K, a carotenoid cation radical is formed having an absorption maximum at 898 nm, an 85 nm blue shift relative to the β-carotene radical cation peak in the WT, and consistent with the formation of the cation radical of a carotenoid with 9 conjugated double bonds. The ratio of Chl+/Car+ is higher in the mutant core complexes, consistent with the higher reduction potential for Car+. As the temperature increases, other carotenoids become accessible to oxidation by P680 +.

Original languageEnglish
Pages (from-to)38839-38850
Number of pages12
JournalJournal of Biological Chemistry
Volume280
Issue number46
DOIs
Publication statusPublished - 2005

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Synechocystis
Photosystem II Protein Complex
Carotenoids
Cations
Genes
Chlorophyll
Railroad cars
Electrons
Rhodobacter capsulatus
Oxidation
Temperature
Near infrared spectroscopy
Near-Infrared Spectroscopy
Oxidation-Reduction
Tyrosine

ASJC Scopus subject areas

  • Biochemistry

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Construction and characterization of genetically modified Synechocystis sp. PCC 6803 photosystem II core complexes containing carotenoids with shorter π-conjugation than β-carotene. / Bautista, James A.; Tracewell, Cara A.; Schlodder, Eberhard; Cunningham, Francis X.; Brudvig, Gary W; Diner, Bruce A.

In: Journal of Biological Chemistry, Vol. 280, No. 46, 2005, p. 38839-38850.

Research output: Contribution to journalArticle

Bautista, James A. ; Tracewell, Cara A. ; Schlodder, Eberhard ; Cunningham, Francis X. ; Brudvig, Gary W ; Diner, Bruce A. / Construction and characterization of genetically modified Synechocystis sp. PCC 6803 photosystem II core complexes containing carotenoids with shorter π-conjugation than β-carotene. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 46. pp. 38839-38850.
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abstract = "β-Carotene has been identified as an intermediate in a secondary electron transfer pathway that oxidizes Chlz and cytochrome b 559 in Photosystem II (PS II) when normal tyrosine oxidation is blocked. To test the redox function of carotenoids in this pathway, we replaced the ζ-carotene desaturase gene (zds) or both the zds and phytoene desaturase (pds) genes of Synechocystis sp. PCC 6803 with the phytoene desaturase gene (crtI) of Rhodobacter capsulatus, producing carotenoids with shorter conjugated π-electron systems and higher reduction potentials than β-carotene. The PS II core complexes of both mutant strains contain approximately the same number of chlorophylls and carotenoids as the wild type but have replaced β-carotene (11 double bonds), with neurosporene (9 conjugated double bonds) and β-zeacarotene (9 conjugated double bonds and 1 β-ionylidene ring). The presence of the ring appears necessary for PS II assembly. Visible and near-infrared spectroscopy were used to examine the light-induced formation of chlorophyll and carotenoid radical cations in the mutant PS II core complexes at temperatures from 20 to 160 K. At 20 K, a carotenoid cation radical is formed having an absorption maximum at 898 nm, an 85 nm blue shift relative to the β-carotene radical cation peak in the WT, and consistent with the formation of the cation radical of a carotenoid with 9 conjugated double bonds. The ratio of Chl+/Car+ is higher in the mutant core complexes, consistent with the higher reduction potential for Car+. As the temperature increases, other carotenoids become accessible to oxidation by P680 +.",
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