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TitleA linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 120, Issue 25, Page e2304833120, Year 2023
Publish dateJun 20, 2023
AuthorsZhen Guo Oh / Warren Shou Leong Ang / Cheng Wei Poh / Soak-Kuan Lai / Siu Kwan Sze / Hoi-Yeung Li / Shashi Bhushan / Tobias Wunder / Oliver Mueller-Cajar /
PubMed AbstractThe slow kinetics and poor substrate specificity of the key photosynthetic CO-fixing enzyme Rubisco have prompted the repeated evolution of Rubisco-containing biomolecular condensates known as ...The slow kinetics and poor substrate specificity of the key photosynthetic CO-fixing enzyme Rubisco have prompted the repeated evolution of Rubisco-containing biomolecular condensates known as pyrenoids in the majority of eukaryotic microalgae. Diatoms dominate marine photosynthesis, but the interactions underlying their pyrenoids are unknown. Here, we identify and characterize the Rubisco linker protein PYCO1 from . PYCO1 is a tandem repeat protein containing prion-like domains that localizes to the pyrenoid. It undergoes homotypic liquid-liquid phase separation (LLPS) to form condensates that specifically partition diatom Rubisco. Saturation of PYCO1 condensates with Rubisco greatly reduces the mobility of droplet components. Cryo-electron microscopy and mutagenesis data revealed the sticker motifs required for homotypic and heterotypic phase separation. Our data indicate that the PYCO1-Rubisco network is cross-linked by PYCO1 stickers that oligomerize to bind to the small subunits lining the central solvent channel of the Rubisco holoenzyme. A second sticker motif binds to the large subunit. Pyrenoidal Rubisco condensates are highly diverse and tractable models of functional LLPS.
External linksProc Natl Acad Sci U S A / PubMed:37311001 / PubMed Central
MethodsEM (single particle)
Resolution2.0 - 2.6 Å
Structure data

33887

7yk5

EMDB-33887, PDB-7yk5:
Rubisco from Phaeodactylum tricornutum bound to PYCO1(452-592)
Method: EM (single particle) / Resolution: 2.0 Å

EMDB-35158: Rubisco from Phaeodactylum tricornutum
Method: EM (single particle) / Resolution: 2.0 Å

EMDB-35159: PYCO1(452-592) from Phaeodactylum tricornutum
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-35166: Rubisco from Phaeodactylum tricornutum bound to PYCO1(452-592)
Method: EM (single particle) / Resolution: 2.2 Å

Chemicals

ChemComp-CAP:
2-CARBOXYARABINITOL-1,5-DIPHOSPHATE

Source
  • phaeodactylum tricornutum (Diatom)
KeywordsPHOTOSYNTHESIS / Rubisco / phase separation / rubisco linker protein / condensation / pyrenoid / phaeodactylum tricornutum

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