This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tooley, A. J.
Right arrow Articles by Glazer, A. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tooley, A. J.
Right arrow Articles by Glazer, A. N.

 Previous Article  |  Next Article 

Journal of Bacteriology, September 2002, p. 4666-4671, Vol. 184, No. 17
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.17.4666-4671.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Biosynthesis of the Cyanobacterial Light-Harvesting Polypeptide Phycoerythrocyanin Holo-{alpha} Subunit in a Heterologous Host

Aaron J. Tooley,{dagger} and Alexander N. Glazer*

Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3200

Received 25 March 2002/ Accepted 30 May 2002

The entire pathway for the biosynthesis of the phycobiliviolin-bearing His-tagged holo-{alpha} subunit of the cyanobacterial photosynthetic accessory protein phycoerythrocyanin was reconstituted in Escherichia coli. Cyanobacterial genes encoding enzymes required for the conversion of heme to 3Z-phycocyanobilin, a precursor of phycobiliviolin (namely, heme oxygenase 1 and 3Z-phycocyanobilin:ferredoxin oxidoreductase), were expressed from a plasmid under the control of the hybrid trp-lac (trc) promoter. Genes for the apo-phycoerythrocyanin {alpha} subunit (pecA) and the heterodimeric lyase/isomerase (pecE and pecF), which catalyzes both the covalent attachment of phycocyanobilin and its concurrent isomerization to phycobiliviolin, were expressed from the trc promoter on a second plasmid. Upon induction, recombinant E. coli used endogenous heme to produce holo-PecA with absorbance and fluorescence properties similar to those of the same protein produced in cyanobacteria. About two-thirds of the apo-PecA was converted to holo-PecA. No significant bilin addition took place in a similarly engineered E. coli strain that lacks pecE and pecF. By using immobilized metal affinity chromatography, both apo-PecA and holo-PecA were isolated as ternary complexes with PecE and PecF. The identities of all three components in the ternary complexes were established unambiguously by protein and tryptic peptide analyses performed by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

* Corresponding author. Mailing address: Natural Reserve System, 1111 Franklin Street, 6th Floor, University of California, Oakland, CA 94607-5200. Phone: (510) 987-0143. Fax: (510) 763-2971. E-mail: glazer{at}uclink4.berkeley.edu.

{dagger} Present address: Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143.

Journal of Bacteriology, September 2002, p. 4666-4671, Vol. 184, No. 17
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.17.4666-4671.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Blot, N., Wu, X.-J., Thomas, J.-C., Zhang, J., Garczarek, L., Bohm, S., Tu, J.-M., Zhou, M., Ploscher, M., Eichacker, L., Partensky, F., Scheer, H., Zhao, K.-H. (2009). Phycourobilin in Trichromatic Phycocyanin from Oceanic Cyanobacteria Is Formed Post-translationally by a Phycoerythrobilin Lyase-Isomerase. J. Biol. Chem. 284: 9290-9298 [Abstract] [Full Text]  
  • Zhao, K.-H., Zhang, J., Tu, J.-M., Bohm, S., Ploscher, M., Eichacker, L., Bubenzer, C., Scheer, H., Wang, X., Zhou, M. (2007). Lyase Activities of CpcS- and CpcT-like Proteins from Nostoc PCC7120 and Sequential Reconstitution of Binding Sites of Phycoerythrocyanin and Phycocyanin beta-Subunits. J. Biol. Chem. 282: 34093-34103 [Abstract] [Full Text]  
  • Shen, G., Saunee, N. A., Williams, S. R., Gallo, E. F., Schluchter, W. M., Bryant, D. A. (2006). Identification and Characterization of a New Class of Bilin Lyase: THE cpcT GENE ENCODES A BILIN LYASE RESPONSIBLE FOR ATTACHMENT OF PHYCOCYANOBILIN TO CYS-153 ON THE beta-SUBUNIT OF PHYCOCYANIN IN SYNECHOCOCCUS SP. PCC 7002. J. Biol. Chem. 281: 17768-17778 [Abstract] [Full Text]  
  • Giepmans, B. N. G., Adams, S. R., Ellisman, M. H., Tsien, R. Y. (2006). The fluorescent toolbox for assessing protein location and function.. Science 312: 217-224 [Abstract] [Full Text]  
  • Zhao, K.-H., Su, P., Li, J., Tu, J.-M., Zhou, M., Bubenzer, C., Scheer, H. (2006). Chromophore Attachment to Phycobiliprotein beta-Subunits: PHYCOCYANOBILIN:CYSTEINE-beta84 PHYCOBILIPROTEIN LYASE ACTIVITY OF CpeS-LIKE PROTEIN FROM ANABAENA Sp. PCC7120. J. Biol. Chem. 281: 8573-8581 [Abstract] [Full Text]  
  • Six, C., Thomas, J.-C., Thion, L., Lemoine, Y., Zal, F., Partensky, F. (2005). Two Novel Phycoerythrin-Associated Linker Proteins in the Marine Cyanobacterium Synechococcus sp. Strain WH8102. J. Bacteriol. 187: 1685-1694 [Abstract] [Full Text]  
  • Frankenberg, N., Lagarias, J. C. (2003). Phycocyanobilin:Ferredoxin Oxidoreductase of Anabaena sp. PCC 7120. BIOCHEMICAL AND SPECTROSCOPIC CHARACTERIZATION. J. Biol. Chem. 278: 9219-9226 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»