Mon1-Ccz1 complex / protein targeting to vacuole / multivesicular body membrane / endosome to lysosome transport via multivesicular body sorting pathway / fungal-type vacuole membrane / autophagy 類似検索 - 分子機能
Vacuolar fusion protein Mon1 / FUZ/MON1/HPS1, third Longin domain / FUZ/MON1/HPS1, second Longin domain / FUZ/MON1/HPS1, first Longin domain / First Longin domain of FUZ, MON1 and HPS1 / Second Longin domain of FUZ, MON1 and HPS1 / Third Longin domain of FUZ, MON1 and HPS1 類似検索 - ドメイン・相同性
ジャーナル: Proc Natl Acad Sci U S A / 年: 2022 タイトル: Structure of the Mon1-Ccz1 complex reveals molecular basis of membrane binding for Rab7 activation. 著者: Björn U Klink / Eric Herrmann / Claudia Antoni / Lars Langemeyer / Stephan Kiontke / Christos Gatsogiannis / Christian Ungermann / Stefan Raunser / Daniel Kümmel / 要旨: Activation of the GTPase Rab7/Ypt7 by its cognate guanine nucleotide exchange factor (GEF) Mon1-Ccz1 marks organelles such as endosomes and autophagosomes for fusion with lysosomes/vacuoles and ...Activation of the GTPase Rab7/Ypt7 by its cognate guanine nucleotide exchange factor (GEF) Mon1-Ccz1 marks organelles such as endosomes and autophagosomes for fusion with lysosomes/vacuoles and degradation of their content. Here, we present a high-resolution cryogenic electron microscopy structure of the Mon1-Ccz1 complex that reveals its architecture in atomic detail. Mon1 and Ccz1 are arranged side by side in a pseudo-twofold symmetrical heterodimer. The three Longin domains of each Mon1 and Ccz1 are triangularly arranged, providing a strong scaffold for the catalytic center of the GEF. At the opposite side of the Ypt7-binding site, a positively charged and relatively flat patch stretches the Longin domains 2/3 of Mon1 and functions as a phosphatidylinositol phosphate-binding site, explaining how the GEF is targeted to membranes. Our work provides molecular insight into the mechanisms of endosomal Rab activation and serves as a blueprint for understanding the function of members of the Tri Longin domain Rab-GEF family.
平均露光時間: 15 sec. / 電子線照射量: 73 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 撮影したグリッド数: 1 / 実像数: 11916 詳細: Images were collected in movie-mode with 4 frames per second
電子光学装置
エネルギーフィルタースリット幅: 20 eV / 位相板: VOLTA PHASE PLATE
画像スキャン
動画フレーム数/画像: 60
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解析
ソフトウェア
名称: PHENIX / バージョン: 1.19.1_4122: / 分類: 精密化
EMソフトウェア
ID
名称
カテゴリ
1
crYOLO
粒子像選択
2
EPU
画像取得
4
MotionCorr2
CTF補正
9
SPHIRE
初期オイラー角割当
10
RELION
最終オイラー角割当
11
RELION
分類
13
Coot
モデル精密化
CTF補正
タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
粒子像の選択
選択した粒子像数: 7155250
3次元再構成
解像度: 3.85 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 911674 / クラス平均像の数: 1 / 対称性のタイプ: POINT