EIPOD fellowship under Marie Sklodowska-Curie Actions COFUND
847543
ドイツ
引用
ジャーナル: Nat Commun / 年: 2024 タイトル: A subgroup of light-driven sodium pumps with an additional Schiff base counterion. 著者: E Podoliak / G H U Lamm / E Marin / A V Schellbach / D A Fedotov / A Stetsenko / M Asido / N Maliar / G Bourenkov / T Balandin / C Baeken / R Astashkin / T R Schneider / A Bateman / J ...著者: E Podoliak / G H U Lamm / E Marin / A V Schellbach / D A Fedotov / A Stetsenko / M Asido / N Maliar / G Bourenkov / T Balandin / C Baeken / R Astashkin / T R Schneider / A Bateman / J Wachtveitl / I Schapiro / V Busskamp / A Guskov / V Gordeliy / A Alekseev / K Kovalev / 要旨: Light-driven sodium pumps (NaRs) are unique ion-transporting microbial rhodopsins. The major group of NaRs is characterized by an NDQ motif and has two aspartic acid residues in the central region ...Light-driven sodium pumps (NaRs) are unique ion-transporting microbial rhodopsins. The major group of NaRs is characterized by an NDQ motif and has two aspartic acid residues in the central region essential for sodium transport. Here we identify a subgroup of the NDQ rhodopsins bearing an additional glutamic acid residue in the close vicinity to the retinal Schiff base. We thoroughly characterize a member of this subgroup, namely the protein ErNaR from Erythrobacter sp. HL-111 and show that the additional glutamic acid results in almost complete loss of pH sensitivity for sodium-pumping activity, which is in contrast to previously studied NaRs. ErNaR is capable of transporting sodium efficiently even at acidic pH levels. X-ray crystallography and single particle cryo-electron microscopy reveal that the additional glutamic acid residue mediates the connection between the other two Schiff base counterions and strongly interacts with the aspartic acid of the characteristic NDQ motif. Hence, it reduces its pKa. Our findings shed light on a subgroup of NaRs and might serve as a basis for their rational optimization for optogenetics.
#262 - 2021年10月 PDB構造への50年間のオープンアクセス (Fifty Years of Open Access to PDB Structures ) 類似性 (3)
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集合体
登録構造単位
A: Bacteriorhodopsin-like protein B: Bacteriorhodopsin-like protein C: Bacteriorhodopsin-like protein D: Bacteriorhodopsin-like protein E: Bacteriorhodopsin-like protein ヘテロ分子
モード: OTHER / 倍率(公称値): 105000 X / 最大 デフォーカス(公称値): 2000 nm / 最小 デフォーカス(公称値): 500 nm / Cs: 2.7 mm / C2レンズ絞り径: 50 µm
撮影
電子線照射量: 50 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 撮影したグリッド数: 1 / 実像数: 3865 / 詳細: Counted super res mode
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解析
EMソフトウェア
ID
名称
バージョン
カテゴリ
1
cryoSPARC
4.0.2
粒子像選択
4
cryoSPARC
4.0.2
CTF補正
10
cryoSPARC
4.0.2
初期オイラー角割当
11
cryoSPARC
4.0.2
最終オイラー角割当
12
cryoSPARC
4.0.2
分類
13
cryoSPARC
4.0.2
3次元再構成
CTF補正
タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
対称性
点対称性: C5 (5回回転対称)
3次元再構成
解像度: 2.63 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 437017 / 対称性のタイプ: POINT
精密化
解像度: 2.63→100.32 Å / Cor.coef. Fo:Fc: 0.581 / SU B: 7.812 / SU ML: 0.152 / ESU R: 0.47 立体化学のターゲット値: MAXIMUM LIKELIHOOD WITH PHASES 詳細: HYDROGENS HAVE BEEN USED IF PRESENT IN THE INPUT