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基本情報
登録情報 | データベース: PDB / ID: 7s9e | ||||||
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タイトル | Cryo-EM Structure of dolphin Prestin: Inhibited II (Sulfate +Salicylate) state | ||||||
![]() | Prestin | ||||||
![]() | MOTOR PROTEIN / Outer hair cells / electromotility / mechanotransduction / hearing / deafness / frequency sensation / echolocation / SLC26 / SLC26A5 | ||||||
機能・相同性 | ![]() cochlear outer hair cell electromotile response / secondary active sulfate transmembrane transporter activity / oxalate transmembrane transporter activity / bicarbonate transmembrane transporter activity / chloride transmembrane transporter activity / sensory perception of sound / regulation of cell shape / plasma membrane 類似検索 - 分子機能 | ||||||
生物種 | ![]() ![]() | ||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.7 Å | ||||||
![]() | Bavi, N. / Clark, M.D. / Contreras, G.F. / Shen, R. / Reddy, B.G. / Milewski, W. / Perozo, E. | ||||||
資金援助 | ![]()
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![]() | ![]() タイトル: The conformational cycle of prestin underlies outer-hair cell electromotility. 著者: Navid Bavi / Michael David Clark / Gustavo F Contreras / Rong Shen / Bharat G Reddy / Wieslawa Milewski / Eduardo Perozo / ![]() 要旨: The voltage-dependent motor protein prestin (also known as SLC26A5) is responsible for the electromotive behaviour of outer-hair cells and underlies the cochlear amplifier. Knockout or impairment of ...The voltage-dependent motor protein prestin (also known as SLC26A5) is responsible for the electromotive behaviour of outer-hair cells and underlies the cochlear amplifier. Knockout or impairment of prestin causes severe hearing loss. Despite the key role of prestin in hearing, the mechanism by which mammalian prestin senses voltage and transduces it into cellular-scale movements (electromotility) is poorly understood. Here we determined the structure of dolphin prestin in six distinct states using single-particle cryo-electron microscopy. Our structural and functional data suggest that prestin adopts a unique and complex set of states, tunable by the identity of bound anions (Cl or SO). Salicylate, a drug that can cause reversible hearing loss, competes for the anion-binding site of prestin, and inhibits its function by immobilizing prestin in a new conformation. Our data suggest that the bound anion together with its coordinating charged residues and helical dipole act as a dynamic voltage sensor. An analysis of all of the anion-dependent conformations reveals how structural rearrangements in the voltage sensor are coupled to conformational transitions at the protein-membrane interface, suggesting a previously undescribed mechanism of area expansion. Visualization of the electromotility cycle of prestin distinguishes the protein from the closely related SLC26 anion transporters, highlighting the basis for evolutionary specialization of the mammalian cochlear amplifier at a high resolution. | ||||||
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構造の表示
ムービー |
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構造ビューア | 分子: ![]() ![]() |
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-関連構造データ
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リンク
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集合体
登録構造単位 | ![]()
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要素
#1: タンパク質 | 分子量: 80973.750 Da / 分子数: 2 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() 遺伝子: SLC26A5 / 発現宿主: ![]() #2: 化合物 | 研究の焦点であるリガンドがあるか | Y | |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: CELL / 3次元再構成法: 単粒子再構成法 |
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試料調製
構成要素 | 名称: Cryo-EM Structure of dolphin Prestin in complex with Salicylate: Inhibited II, Sulfate + Salicylate タイプ: COMPLEX / Entity ID: #1 / 由来: RECOMBINANT |
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由来(天然) | 生物種: ![]() ![]() |
由来(組換発現) | 生物種: ![]() |
緩衝液 | pH: 7.4 詳細: 125 mM Na2SO4, 5mM Mg(OH)2, 20 mM Tris-OH, 10-15 mM methanesulfonic acid + 0.02 % GDN + 50 mM Sodium Salicylate |
緩衝液成分 | 濃度: 125 mM / 名称: sodium sulfate / 式: Na2SO4 |
試料 | 濃度: 2.5 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES 詳細: Monodisperse peak at around 14 ml using SEC (Superose 6 column) |
試料支持 | グリッドのタイプ: Quantifoil R1.2/1.3 |
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 295 K 詳細: 4.5 s blot times, blot force 3, and double filter papers on each side of the vitrobot. |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD |
撮影 | 電子線照射量: 1 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) |
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解析
CTF補正 | タイプ: NONE |
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3次元再構成 | 解像度: 3.7 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 112000 / 対称性のタイプ: POINT |