Engineering and Physical Sciences Research Council
EP/W024063/1
英国
Future Leader Fellowship
MR/T020679/1
英国
Biotechnology and Biological Sciences Research Council (BBSRC)
BB/S015337/1
英国
Biotechnology and Biological Sciences Research Council (BBSRC)
BB/S015337/1
英国
Future Leader Fellowship
MR/T040742/1
英国
引用
ジャーナル: Nat Plants / 年: 2024 タイトル: A promiscuous mechanism to phase separate eukaryotic carbon fixation in the green lineage. 著者: James Barrett / Mihris I S Naduthodi / Yuwei Mao / Clément Dégut / Sabina Musiał / Aidan Salter / Mark C Leake / Michael J Plevin / Alistair J McCormick / James N Blaza / Luke C M Mackinder / 要旨: CO fixation is commonly limited by inefficiency of the CO-fixing enzyme Rubisco. Eukaryotic algae concentrate and fix CO in phase-separated condensates called pyrenoids, which complete up to one- ...CO fixation is commonly limited by inefficiency of the CO-fixing enzyme Rubisco. Eukaryotic algae concentrate and fix CO in phase-separated condensates called pyrenoids, which complete up to one-third of global CO fixation. Condensation of Rubisco in pyrenoids is dependent on interaction with disordered linker proteins that show little conservation between species. We developed a sequence-independent bioinformatic pipeline to identify linker proteins in green algae. We report the linker from Chlorella and demonstrate that it binds a conserved site on the Rubisco large subunit. We show that the Chlorella linker phase separates Chlamydomonas Rubisco and that despite their separation by ~800 million years of evolution, the Chlorella linker can support the formation of a functional pyrenoid in Chlamydomonas. This cross-species reactivity extends to plants, with the Chlorella linker able to drive condensation of some native plant Rubiscos in vitro and in planta. Our results represent an exciting frontier for pyrenoid engineering in plants, which is modelled to increase crop yields.
モデル: Quantifoil R1.2/1.3 / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 支持フィルム - Film thickness: 11 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 60 sec. / 前処理 - 雰囲気: OTHER / 詳細: The pressure was not recorded
凍結
凍結剤: ETHANE / チャンバー内湿度: 95 % / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK IV / 詳細: Blot force -5, 8 s blot.
詳細
Chlorella Rubisco at 0.3 mg/mL (0.5 micromolar) was mixed with the alpha3-alpha4 fragment at 16 micromolar and incubated for 10 minutes at room temperature before grid preparation.
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電子顕微鏡法
顕微鏡
TFS GLACIOS
撮影
フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) 撮影したグリッド数: 1 / 実像数: 4447 / 平均露光時間: 6.52 sec. / 平均電子線量: 50.0 e/Å2