- EMDB-20914: Human Connexin-26 (Neutral pH open conformation) -
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基本情報
登録情報
データベース: EMDB / ID: EMD-20914
タイトル
Human Connexin-26 (Neutral pH open conformation)
マップデータ
Sharpened map at 4.0 A resolution.
試料
細胞器官・細胞要素: Connexin-26 Gap Junction Channel
タンパク質・ペプチド: Gap junction beta-2 protein
機能・相同性
機能・相同性情報
Transport of connexons to the plasma membrane / response to human chorionic gonadotropin / gap junction-mediated intercellular transport / epididymis development / gap junction channel activity involved in cell communication by electrical coupling / Oligomerization of connexins into connexons / Transport of connexins along the secretory pathway / gap junction assembly / connexin complex / Gap junction assembly ...Transport of connexons to the plasma membrane / response to human chorionic gonadotropin / gap junction-mediated intercellular transport / epididymis development / gap junction channel activity involved in cell communication by electrical coupling / Oligomerization of connexins into connexons / Transport of connexins along the secretory pathway / gap junction assembly / connexin complex / Gap junction assembly / gap junction / astrocyte projection / gap junction channel activity / cellular response to glucagon stimulus / inner ear development / decidualization / endoplasmic reticulum-Golgi intermediate compartment / lateral plasma membrane / response to retinoic acid / cellular response to dexamethasone stimulus / response to ischemia / response to progesterone / sensory perception of sound / transmembrane transport / cell-cell signaling / response to estradiol / cellular response to oxidative stress / cell body / response to lipopolysaccharide / calcium ion binding / perinuclear region of cytoplasm / identical protein binding / plasma membrane 類似検索 - 分子機能
Gap junction beta-2 protein (Cx26) / Connexin / Connexin, N-terminal / Connexin, conserved site / Gap junction protein, cysteine-rich domain / Connexin, N-terminal domain superfamily / Connexin / Connexins signature 1. / Connexins signature 2. / Connexin homologues / Gap junction channel protein cysteine-rich domain 類似検索 - ドメイン・相同性
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01 HL48908
米国
National Institutes of Health/National Human Genome Research Institute (NIH/NHGRI)
F30 HL129702-01
米国
引用
ジャーナル: Cell Rep / 年: 2020 タイトル: A Steric "Ball-and-Chain" Mechanism for pH-Mediated Regulation of Gap Junction Channels. 著者: Ali K Khan / Maciej Jagielnicki / William E McIntire / Michael D Purdy / Venkatasubramanian Dharmarajan / Patrick R Griffin / Mark Yeager / 要旨: Gap junction channels (GJCs) mediate intercellular communication and are gated by numerous conditions such as pH. The electron cryomicroscopy (cryo-EM) structure of Cx26 GJC at physiological pH ...Gap junction channels (GJCs) mediate intercellular communication and are gated by numerous conditions such as pH. The electron cryomicroscopy (cryo-EM) structure of Cx26 GJC at physiological pH recapitulates previous GJC structures in lipid bilayers. At pH 6.4, we identify two conformational states, one resembling the open physiological-pH structure and a closed conformation that displays six threads of density, that join to form a pore-occluding density. Crosslinking and hydrogen-deuterium exchange mass spectrometry reveal closer association between the N-terminal (NT) domains and the cytoplasmic loops (CL) at acidic pH. Previous electrophysiologic studies suggest an association between NT residue N14 and H100 near M2, which may trigger the observed movement of M2 toward M1 in our cryo-EM maps, thereby accounting for additional NT-CL crosslinks at acidic pH. We propose that these pH-induced interactions and conformational changes result in extension, ordering, and association of the acetylated NT domains to form a hexameric "ball-and-chain" gating particle.
詳細: NaCl was diluted from 1 M to 200 mM immediately before freezing.
グリッド
モデル: C-flat-2/2
凍結
凍結剤: ETHANE / 装置: HOMEMADE PLUNGER 詳細: After application of sample to the C-flat holey carbon grid, sample was manually blotted with Whatman no. 1 filter paper for 3-5 s..
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
特殊光学系
エネルギーフィルター - 名称: GIF Quantum LS / エネルギーフィルター - スリット幅: 20 eV
撮影
フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / 撮影したグリッド数: 1 / 平均露光時間: 8.0 sec. / 平均電子線量: 45.0 e/Å2
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 倍率(公称値): 130000
実験機器
モデル: Titan Krios / 画像提供: FEI Company
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画像解析
粒子像選択
選択した数: 64087
CTF補正
ソフトウェア: (名称: CTFFIND (ver. 4), RELION)
初期モデル
モデルのタイプ: OTHER 詳細: 2D class averages of the auto-picked particles were calculated and used to generate a de novo D6 symmetric initial model in EMAN2.
Initial local fitting was done using UCSF Chimera. Multiple rounds of automated (Phenix) and manual (Coot) model building were performed. The model was validated in Molprobity(part of Phenix package). Model-map cross correlation score as well as EMRinger score were obtained in Phenix. RMSD values for C-alphas were calculated in UCSF Chimera.
精密化
空間: REAL / プロトコル: OTHER / 当てはまり具合の基準: Model-map cross correlation
得られたモデル
PDB-6uvr: Human Connexin-26 (Neutral pH open conformation)