EMDB-28740: Brain-derived 42-residue amyloid-beta fibril type A

基本情報

登録情報

データベース: EMDB / ID: EMD-28740

• タイトル: Brain-derived 42-residue amyloid-beta fibril type A

試料

• 複合体: amyloid-b 42 (Ab42) fibril
  • タンパク質・ペプチド: Beta-amyloid protein 42アミロイドβ

機能・相同性

分子機能:

regulation of epidermal growth factor-activated receptor activity / signaling receptor activator activity / cytosolic mRNA polyadenylation / collateral sprouting in absence of injury / microglia development / regulation of synapse structure or activity / Formyl peptide receptors bind formyl peptides and many other ligands / regulation of Wnt signaling pathway / axo-dendritic transport / synaptic assembly at neuromuscular junction / smooth endoplasmic reticulum calcium ion homeostasis / axon midline choice point recognition / astrocyte activation involved in immune response / regulation of spontaneous synaptic transmission / mating behavior / NMDA selective glutamate receptor signaling pathway / 繊毛 / Lysosome Vesicle Biogenesis / PTB domain binding / Golgi-associated vesicle / positive regulation of amyloid fibril formation / neuron remodeling / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / protein serine/threonine kinase binding / Deregulated CDK5 triggers multiple neurodegenerative pathways in Alzheimer's disease models / : / nuclear envelope lumen / suckling behavior / presynaptic active zone / dendrite development / COPII-coated ER to Golgi transport vesicle / modulation of excitatory postsynaptic potential / TRAF6 mediated NF-kB activation / Advanced glycosylation endproduct receptor signaling / neuromuscular process controlling balance / The NLRP3 inflammasome / regulation of presynapse assembly / transition metal ion binding / regulation of multicellular organism growth / intracellular copper ion homeostasis / negative regulation of long-term synaptic potentiation / negative regulation of neuron differentiation / ECM proteoglycans / 小胞体 / Mitochondrial protein degradation / positive regulation of T cell migration / spindle midzone / Purinergic signaling in leishmaniasis infection / positive regulation of calcium-mediated signaling / クラスリン / regulation of peptidyl-tyrosine phosphorylation / positive regulation of chemokine production / forebrain development / Notchシグナリング / positive regulation of G2/M transition of mitotic cell cycle / neuron projection maintenance / positive regulation of protein metabolic process / ionotropic glutamate receptor signaling pathway / positive regulation of glycolytic process / cholesterol metabolic process / response to interleukin-1 / positive regulation of mitotic cell cycle / extracellular matrix organization / 軸索誘導 / adult locomotory behavior / trans-Golgi network membrane / platelet alpha granule lumen / locomotory behavior / positive regulation of peptidyl-threonine phosphorylation / dendritic shaft / 学習 / positive regulation of interleukin-1 beta production / central nervous system development / positive regulation of long-term synaptic potentiation / endosome lumen / astrocyte activation / Post-translational protein phosphorylation / positive regulation of JNK cascade / synapse organization / regulation of long-term neuronal synaptic plasticity / microglial cell activation / TAK1-dependent IKK and NF-kappa-B activation / visual learning / serine-type endopeptidase inhibitor activity / neuromuscular junction / recycling endosome / 認識 / positive regulation of inflammatory response / Golgi lumen / neuron cellular homeostasis / エンドサイトーシス / positive regulation of non-canonical NF-kappaB signal transduction / cellular response to amyloid-beta / positive regulation of interleukin-6 production / Regulation of Insulin-like Growth Factor (IGF) transport and uptake by Insulin-like Growth Factor Binding Proteins (IGFBPs) / neuron projection development / G2/M transition of mitotic cell cycle / positive regulation of tumor necrosis factor production / cell-cell junction / シナプス小胞

ドメイン・相同性:

Amyloidogenic glycoprotein, copper-binding / Amyloidogenic glycoprotein, copper-binding domain conserved site / Amyloidogenic glycoprotein, copper-binding domain superfamily / Copper-binding of amyloid precursor, CuBD / Amyloid precursor protein (APP) copper-binding (CuBD) domain signature. / Amyloidogenic glycoprotein, amyloid-beta peptide superfamily / Beta-amyloid peptide (beta-APP) / Amyloidogenic glycoprotein, amyloid-beta peptide / Beta-amyloid precursor protein C-terminal / Amyloidogenic glycoprotein, intracellular domain, conserved site / Beta-amyloid precursor protein C-terminus / Amyloid precursor protein (APP) intracellular domain signature. / Amyloid precursor protein (APP) E1 domain profile. / Amyloid precursor protein (APP) E2 domain profile. / Amyloidogenic glycoprotein, extracellular / Amyloidogenic glycoprotein, heparin-binding / Amyloidogenic glycoprotein, E2 domain / E2 domain superfamily / Amyloidogenic glycoprotein, heparin-binding domain superfamily / Amyloid A4 N-terminal heparin-binding / E2 domain of amyloid precursor protein / amyloid A4 / Amyloidogenic glycoprotein / Proteinase inhibitor I2, Kunitz, conserved site / Pancreatic trypsin inhibitor (Kunitz) family signature. / BPTI/Kunitz family of serine protease inhibitors. / Pancreatic trypsin inhibitor Kunitz domain / Kunitz/Bovine pancreatic trypsin inhibitor domain / Pancreatic trypsin inhibitor (Kunitz) family profile. / Pancreatic trypsin inhibitor Kunitz domain superfamily / PH-like domain superfamily

構成要素:

アミロイド前駆体タンパク質

• 生物種: Homo sapiens (ヒト)
• 手法: らせん対称体再構成法 / クライオ電子顕微鏡法 / 解像度: 2.83 Å
• データ登録者: Tycko R / Lee M / Yau Y-M / Louis JM

資金援助

米国, 1件

Organization	Grant number	国
Not funded		米国

• 引用: ジャーナル: Proc Natl Acad Sci U S A / 年: 2023; タイトル: Structures of brain-derived 42-residue amyloid-β fibril polymorphs with unusual molecular conformations and intermolecular interactions.; 著者: Myungwoon Lee / Wai-Ming Yau / John M Louis / Robert Tycko / ; 要旨: Fibrils formed by the 42-residue amyloid-β peptide (Aβ42), a main component of amyloid deposits in Alzheimer's disease (AD), are known to be polymorphic, i.e., to contain multiple possible molecular structures. Previous studies of Aβ42 fibrils, including fibrils prepared entirely in vitro or extracted from brain tissue and using solid-state NMR (ssNMR) or cryogenic electron microscopy (cryo-EM) methods, have found polymorphs with differences in amino acid sidechain orientations, lengths of structurally ordered segments, and contacts between cross-β subunit pairs within a single filament. Despite these differences, Aβ42 molecules adopt a common S-shaped conformation in all previously described high-resolution Aβ42 fibril structures. Here we report two cryo-EM-based structures of Aβ42 fibrils that are qualitatively different, in samples derived from AD brain tissue by seeded growth. In type A fibrils, residues 12 to 42 adopt a ν-shaped conformation, with both intra-subunit and intersubunit hydrophobic contacts to form a compact core. In type B fibrils, residues 2 to 42 adopt an υ-shaped conformation, with only intersubunit contacts and internal pores. Type A and type B fibrils have opposite helical handedness. Cryo-EM density maps and molecular dynamics simulations indicate intersubunit K16-A42 salt bridges in type B fibrils and partially occupied K28-A42 salt bridges in type A fibrils. The coexistence of two predominant polymorphs, with differences in N-terminal dynamics, is supported by ssNMR data, as is faithful propagation of structures from first-generation to second-generation brain-seeded Aβ42 fibril samples. These results demonstrate that Aβ42 fibrils can exhibit a greater range of structural variations than seen in previous studies.

履歴

登録	2022年10月31日	-
ヘッダ(付随情報) 公開	2023年3月22日	-
マップ公開	2023年3月22日	-
更新	2023年3月22日	-
現状	2023年3月22日	処理サイト: RCSB / 状態: 公開

マップ

• ファイル: ダウンロード / ファイル: emd_28740.map.gz / 形式: CCP4 / 大きさ: 476.8 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• ボクセルのサイズ: X=Y=Z: 0.86 Å

密度

表面レベル	登録者による: 0.009
最小 - 最大	-0.037047435 - 0.08876042
平均 (標準偏差)	9.7738e-06 (±0.0015273857)

• 対称性: 空間群: 1

詳細

EMDB XML:

マップ形状	Axis order X Y Z Origin 0 0 0 サイズ 500 500 500 Spacing 500 500 500
セル	A=B=C: 430.0 Å α=β=γ: 90.0 °

添付データ

ハーフマップ: #1

• ファイル: emd_28740_half_map_1.map

ハーフマップ: #2

• ファイル: emd_28740_half_map_2.map

試料の構成要素

全体 : amyloid-b 42 (Ab42) fibril

• 全体: 名称: amyloid-b 42 (Ab42) fibril

要素

• 複合体: amyloid-b 42 (Ab42) fibril
  • タンパク質・ペプチド: Beta-amyloid protein 42アミロイドβ

超分子 #1: amyloid-b 42 (Ab42) fibril

• 超分子: 名称: amyloid-b 42 (Ab42) fibril / タイプ: complex / ID: 1 / キメラ: Yes / 親要素: 0 / 含まれる分子: all
• 由来(天然): 生物種: Homo sapiens (ヒト)
• 分子量: 理論値: 4514.10 kDa/nm

分子 #1: Beta-amyloid protein 42

• 分子: 名称: Beta-amyloid protein 42 / タイプ: protein_or_peptide / ID: 1 / コピー数: 8 / 光学異性体: LEVO
• 由来(天然): 生物種: Homo sapiens (ヒト)
• 分子量: 理論値: 4.520087 KDa
• 組換発現: 生物種: Escherichia coli BL21(DE3) (大腸菌)

配列

文字列:

DAEFRHDSGY EVHHQKLVFF AEDVGSNKGA IIGLMVGGVV IA

実験情報

構造解析

• 手法: クライオ電子顕微鏡法
• 解析: らせん対称体再構成法
• 試料の集合状態: filament

試料調製

• 濃度: 0.34 mg/mL
• 緩衝液: pH: 7.4 / 構成要素 - 濃度: 10.0 mM / 構成要素 - 名称: Sodium Phosphate / 詳細: 10mM Na-phosphate, 0.1% sodium azide
• 凍結: 凍結剤: ETHANE / チャンバー内湿度: 99 % / チャンバー内温度: 93 K / 装置: FEI VITROBOT MARK I; 詳細: Preblot for 12-13 seconds and blot for 2.5-3.0 seconds before plunging.

電子顕微鏡法

• 顕微鏡: FEI TITAN KRIOS
• 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: FLOOD BEAM / 撮影モード: BRIGHT FIELDBright-field microscopy / 最大 デフォーカス（公称値）: 3.0 µm / 最小 デフォーカス（公称値）: 0.5 µm / 倍率（公称値）: 130000
• 試料ステージ: 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER; ホルダー冷却材: NITROGEN
• 撮影: フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k); 検出モード: SUPER-RESOLUTION / デジタル化 - サイズ - 横: 11520 pixel / デジタル化 - サイズ - 縦: 8184 pixel / 撮影したグリッド数: 1 / 実像数: 3383 / 平均露光時間: 1.65 sec. / 平均電子線量: 44.65 e/Ų
• 実験機器: モデル: Titan Krios / 画像提供: FEI Company

画像解析

• Segment selection: 選択した数: 374821 / ソフトウェア - 名称: RELION (ver. 4.0.0)
• 初期モデル: モデルのタイプ: NONE / 詳細: featureless cylinder
• 最終 角度割当: タイプ: NOT APPLICABLE / ソフトウェア - 名称: RELION (ver. 4.0.0); 詳細: 2.46 Angstrom helical rise and -179.09 degree helical twist
• 最終 再構成: 想定した対称性 - らせんパラメータ - Δz: 2.46 Å; 想定した対称性 - らせんパラメータ - ΔΦ: -179.09 °; 想定した対称性 - らせんパラメータ - 軸対称性: C₁ (非対称); 解像度のタイプ: BY AUTHOR / 解像度: 2.83 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: RELION (ver. 4.0.0) / 使用した粒子像数: 68481
• 詳細: Gatan Imaging Filter (GIF) Quantum LS

原子モデル構築 1

• 詳細: Manually generated model was fit into the density using PHENIX and UCSF Chimera. Further refinements were performed using Xplor-NIH.
• 精密化: プロトコル: OTHER

得られたモデル

PDB-8ezd:
Brain-derived 42-residue amyloid-beta fibril type A