Analysis of Carboxysomes from Synechococcus PCC7942 Reveals Multiple Rubisco Complexes with Carboxysomal Proteins CcmM and CcaA

• Published in: The Journal of biological chemistry Vol. 282; no. 40; pp. 29323 - 29335
• Main Authors: Long, Benedict M., Badger, Murray R., Whitney, Spencer M., Price, G. Dean
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05-10-2007; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Bacterial Proteins - chemistry; Base Sequence; Chromatography - methods; Cyanobacteria - metabolism; Escherichia coli - metabolism; Genetic Complementation Test; Mass Spectrometry; Molecular Sequence Data; Peptides - chemistry; Protein Binding; Proteomics - methods; Ribulose-Bisphosphate Carboxylase - chemistry; Ribulose-Bisphosphate Carboxylase - metabolism; Sequence Homology, Amino Acid; Synechococcus; Synechococcus - genetics; Synechococcus - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M703896200

In cyanobacteria, the key enzyme for photosynthetic CO2 fixation, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), is bound within proteinaceous polyhedral microcompartments called carboxysomes. Cyanobacteria with Form IB Rubisco produce β-carboxysomes whose putative shell proteins are encoded by the ccm-type genes. To date, very little is known of the protein-protein interactions that form the basis of β-carboxysome structure. In an effort to identify such interactions within the carboxysomes of the β-cyanobacterium Synechococcus sp. PCC7942, we have used polyhistidine-tagging approaches to identify at least three carboxysomal subcomplexes that contain active Rubisco. In addition to the expected L8S8 Rubisco, which is the major component of carboxysomes, we have identified two Rubisco complexes containing the putative shell protein CcmM, one of which also contains the carboxysomal carbonic anhydrase, CcaA. The complex containing CcaA consists of Rubisco and the full-length 58-kDa form of CcmM (M58), whereas the other is made up of Rubisco and a short 35-kDa form of CcmM (M35), which is probably translated independently of M58 via an internal ribosomal entry site within the ccmM gene. We also show that the high CO2-requiring ccmM deletion mutant (ΔccmM) can achieve nearly normal growth rates at ambient CO2 after complementation with both wild type and chimeric (His6-tagged) forms of CcmM. Although a significant amount of independent L8S8 Rubisco is confined to the center of the carboxysome, we speculate that the CcmM-CcaA-Rubisco complex forms an important assembly coordination within the carboxysome shell. A speculative carboxysome structural model is presented.