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TitleUnique biogenesis and kinetics of hornwort Rubiscos revealed by synthetic biology systems.
Journal, issue, pagesMol Plant, Year 2024
Publish dateNov 2, 2024
AuthorsZhen-Guo Oh / Tanner Ashton Robison / Dan-Hong Loh / Warren Shou Leong Ang / Jediael Ng / Fay-Wei Li / Laura Helen Gunn /
PubMed AbstractHornworts are the only land plants that employ a pyrenoid to optimize Rubisco's CO fixation. Yet, hornwort Rubisco remains poorly characterized. Here we assemble the hornwort Anthoceros agrestis ...Hornworts are the only land plants that employ a pyrenoid to optimize Rubisco's CO fixation. Yet, hornwort Rubisco remains poorly characterized. Here we assemble the hornwort Anthoceros agrestis Rubisco (AaRubisco) using the Arabidopsis thaliana SynBio expression system and observed the formation of stalled intermediates, prompting us to develop a new SynBio system with A. agrestis cognate chaperones. We successfully assembled AaRubisco and Rubisco from three other hornwort species. Unlike A. thaliana Rubisco, AaRubisco assembly is not dependent on RbcX or Raf2. Kinetic characterization reveals that hornwort Rubiscos exhibit a range of catalytic rates (3-10 s), but with similar affinity (∼30 μM) and specificity (∼70) for CO. In other words, hornwort Rubiscos do not comply with the long held canonical catalytic trade-off observed in other land plants, providing experimental support that Rubisco kinetics may be phylogenetically constrained. Unexpectedly, we observed a 50% increase in AaRubisco catalytic rates when RbcX was removed from our SynBio system, without any reduction in specificity. Structural biology, biochemistry and proteomic analysis suggest that subtle differences in Rubisco large subunit interactions, when RbcX is absent during biogenesis, increases the accessibility of active sites and catalytic turnover rate. This study uncovered a previously unknown Rubisco kinetic parameter space and provides a SynBio chassis to expand the survey of other Rubisco kinetics. Our discovery could thus reshape the approaches for engineering Rubisco with superior kinetics.
External linksMol Plant / PubMed:39491367
MethodsEM (single particle)
Resolution2.88 - 3.0 Å
Structure data

45605

9chz

EMDB-45605, PDB-9chz:
Anthoceros agrestis Rubisco assembled with Raf1 Raf2 and BSD2
Method: EM (single particle) / Resolution: 2.9 Å

45607

9ci1

EMDB-45607, PDB-9ci1:
Anthoceros agrestis Rubisco octamer core complexed with Arabidopsis thaliana BSD2
Method: EM (single particle) / Resolution: 2.88 Å

45608

9ci2

EMDB-45608, PDB-9ci2:
Anthoceros agrestis Rubisco octamer core complexed with small subunits and Arabidopsis thaliana BSD2
Method: EM (single particle) / Resolution: 2.9 Å

45641

9ck5

EMDB-45641, PDB-9ck5:
Anthoceros agrestis Rubisco assembled with RbcX1, RbcX2, Raf1, Raf2 and BSD2
Method: EM (single particle) / Resolution: 3.0 Å

Chemicals

ChemComp-MG:
Unknown entry

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

Source
  • anthoceros agrestis (plant)
  • arabidopsis thaliana (thale cress)
KeywordsPHOTOSYNTHESIS / Anthoceros agrestis rubisco

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