Structure and assembly of cargo Rubisco in two native α-carboxysomes

• Published in: Nature communications 2022-07, Vol.13 (1), p.4299-9, Article 4299
• Main Authors: Ni, Tao, Sun, Yaqi, Burn, Will, Al-Hazeem, Monsour M. J., Zhu, Yanan, Yu, Xiulian, Liu, Lu-Ning, Zhang, Peijun
• Format: Article
• Language: English
• Subjects: 101/28; 631/326; 631/449/1734/1790; 631/535/1258/1260; Assembly; Bacterial Proteins - metabolism; Carbon; Carbon Cycle; Carbon Dioxide - metabolism; Carbon fixation; Carbonic anhydrase; Carbonic anhydrases; Cargo ships; Crystal structure; Cyanobacteria; Cyanobacteria - metabolism; Cyanobium; Encapsulation; Enzymes; Fixation; Halothiobacillus - genetics; Halothiobacillus - metabolism; Humanities and Social Sciences; Intrinsically Disordered Proteins - metabolism; Metabolism; multidisciplinary; Organelles - metabolism; Oxygenase; Peptides; Proteins; Ribulose-bisphosphate carboxylase; Ribulose-Bisphosphate Carboxylase - metabolism; Science; Science (multidisciplinary); Spirals; Tomography
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-32004-w

Carboxysomes are a family of bacterial microcompartments in cyanobacteria and chemoautotrophs. They encapsulate Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and carbonic anhydrase catalyzing carbon fixation inside a proteinaceous shell. How Rubisco complexes pack within the carboxysomes is unknown. Using cryo-electron tomography, we determine the distinct 3D organization of Rubisco inside two distant α-carboxysomes from a marine α-cyanobacterium Cyanobium sp. PCC 7001 where Rubiscos are organized in three concentric layers, and from a chemoautotrophic bacterium Halothiobacillus neapolitanus where they form intertwining spirals. We further resolve the structures of native Rubisco as well as its higher-order assembly at near-atomic resolutions by subtomogram averaging. The structures surprisingly reveal that the authentic intrinsically disordered linker protein CsoS2 interacts with Rubiscos in native carboxysomes but functions distinctively in the two α-carboxysomes. In contrast to the uniform Rubisco-CsoS2 association in the Cyanobium α-carboxysome, CsoS2 binds only to the Rubiscos close to the shell in the Halo α-carboxysome. Our findings provide critical knowledge of the assembly principles of α-carboxysomes, which may aid in the rational design and repurposing of carboxysome structures for new functions. Carboxysomes are bacterial microcompartments encapsulating Rubisco and carbonic anhydrase for carbon fixation. Here, authors determine the organization of Rubisco and its interaction with the linker protein CsoS2 within two distant α-carboxysomes.