13C magic angle spinning NMR evidence for a 15,15'-cis configuration of the spheroidene in the Rhodobacter sphaeroides photosynthetic reaction center.
The photosynthetic reaction center of Rhodobacter sphaeroides 2.4.1 contains one carotenoid that protects the protein complex against photodestruction. The structure around the central (15,15') double bond of the bound spheroidene carotenoid was investigated with low-temperature magic angle spinning 13C NMR, which allows an in situ characterization of the configuration of the central double bond in the carotenoid. Carotenoidless reaction centers of R. sphaeroides R26 were reconstituted with spheroidene specifically labeled at the C-14' or C-15' position, and the signals from the labels were separated from the natural abundance background using 13C MAS NMR difference spectroscopy. The resonances shift 5.2 and 3.8 ppm upfield upon incorporation in the protein complex, similar to the 5.6 and 4.4 ppm upfield shift occurring in the model compound beta-carotene upon trans to 15,15'-cis isomerization. Hence the MAS NMR favors a cis configuration, as opposed to the trans configuration deduced from X-ray data.
de Groot
H J
HJ
Department of Chemistry, Gorlaeus Laboratories, Leiden, The Netherlands.
Gebhard
R
R
van der Hoef
I
I
Hoff
A J
AJ
Lugtenburg
J
J
Violette
C A
CA
Frank
H A
HA
D002247:Carbon Isotopes; D002338:Carotenoids / Q000032:analysis / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008968:Molecular Conformation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D012242:Rhodobacter sphaeroides / Q000737:chemistry*
D002247:Carbon Isotopes; D045322:Photosynthetic Reaction Center Complex Proteins; C005637:spheroidene; D002338:Carotenoids