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TitleStructure of Chlamydomonas reinhardtii LciA guided the engineering of FNT family proteins to gain bicarbonate transport activity.
Journal, issue, pagesNat Plants, Vol. 12, Issue 1, Page 231-240, Year 2026
Publish dateJan 8, 2026
AuthorsJiaxin Guo / Zhao Yang / Xue Zhang / Feifan Liu / Miaolian Ma / Fang Yu / Jirong Huang / Peng Zhang /
PubMed AbstractEngineering functional CO-concentrating mechanisms into C crops holds great potential for enhancing photosynthetic efficiency. Limited CO-inducible A (LciA), a chloroplast envelope bicarbonate ...Engineering functional CO-concentrating mechanisms into C crops holds great potential for enhancing photosynthetic efficiency. Limited CO-inducible A (LciA), a chloroplast envelope bicarbonate channel belonging to the formate/nitrite transporter (FNT) family, is a key algal CO2-concentrating mechanism component and has been considered as a prime candidate for introduction into C plants. However, its application has been hindered by an incomplete mechanistic understanding. Here we report the cryogenic electron microscopy structure of Chlamydomonas reinhardtii LciA. Combining structural analysis and growth assays, we determined key residues governing substrate access and permeation, and identified two substitutions (K136A/A114F) that enhance LciA activity. We found that bicarbonate selectivity is governed by electrostatic coordination mediated by Lys220 and steric constraint imposed by Ala117 and Val267 within the selectivity filter. Leveraging these insights, we successfully engineered the bacterial FNT family nitrite channel NirC through site-directed mutagenesis to gain bicarbonate transport activity, and we characterized the bicarbonate transport capacity of the Chlamydomonas nitrite channels NAR1.1/NAR1.5, which were amenable to further enhancement. Taken together, our study establishes LciA as a fundamental template for engineering and identifying FNT proteins with bicarbonate transport capability, thereby greatly expanding the molecular toolkit for synthetic biology approaches aimed at boosting photosynthetic efficiency in both algae and crops.
External linksNat Plants / PubMed:41507353
MethodsEM (single particle)
Resolution2.37 Å
Structure data

EMDB-64722, PDB-9v2a:
The Chlamydomonas reinhardtii bicarbonate transporter LciA
Method: EM (single particle) / Resolution: 2.37 Å

Source
  • chlamydomonas reinhardtii (plant)
KeywordsTRANSPORT PROTEIN / Chlamydomonas reinhardtii / bicarbonate transporters / LciA / CO2-concentrating mechanism

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