Abstract
The phycobilin: Cysteine-84-phycobiliprotein lyase, CpeS1, catalyzes phycocyanobilin (PCB) and phycoerythrobilin attachment to nearly all cysteine-84 (consensus sequence) binding sites of phycoerythrin, phycoerythrocyanin, phycocyanin and allophycocyanin (Zhao et al. (2007) Proc Natl Acad Sci 104:14300–14305). We now show that CpeS1 can bind PCB, as assayed by Ni2+ chelating affinity chromatography. Binding is rapid, and the chromophore is bound in an extended conformation similar to that in phycobiliproteins but only poorly fluorescent. Upon addition of apo-biliproteins, the chromophore is transferred to the latter much slower (∼1 h), indicating that chromophorylated CpeS1 is an intermediate in the enzymatic reaction. In addition, imidazole is bound to PCB, as shown by mass spectroscopy of tryptic digests of the intermediate CpeS1–PCB complex.
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Abbreviations
- CPC:
-
Phycocyanin
- CpcB:
-
Apo-β-CPC
- CpeS1:
-
Phycobilin
- Cys84:
-
Phycobiliprotein lyase
- KPB:
-
Potassium phosphate buffer
- PCB:
-
Phycocyanobilin
- PEC:
-
Phycoerythrocyanin
- PecB:
-
Apo-β-PEC
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Acknowledgments
HS and KHZ are grateful to Volkswagen Stiftung for the Partnership (I/77900). HS is grateful to Deutsche Forschungsgemeinschaft for the grant (SFB 533, TPA1), KHZ to Program for New Century Excellent Talents in University for the grant (NCET-04-0717, P.R. China), and JMT to Chinese Scholarship Council for a doctoral scholarship.
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Tu, JM., Kupka, M., Böhm, S. et al. Intermediate binding of phycocyanobilin to the lyase, CpeS1, and transfer to apoprotein. Photosynth Res 95, 163–168 (2008). https://doi.org/10.1007/s11120-007-9251-4
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DOI: https://doi.org/10.1007/s11120-007-9251-4