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データを開く
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基本情報
登録情報 | データベース: PDB / ID: 7sch | ||||||
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タイトル | Cryo-EM structure of the human Exostosin-1 and Exostosin-2 heterodimer | ||||||
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![]() | TRANSFERASE / exostosin1 / exostosin2 / glycosyltransferase / heparan sulfate | ||||||
機能・相同性 | ![]() glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase / hypersensitivity / heart field specification / lymphocyte adhesion to endothelial cell of high endothelial venule / heparan sulfate N-acetylglucosaminyltransferase activity / glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase activity / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase activity / smoothened signaling pathway involved in lung development / developmental growth involved in morphogenesis ...glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase / hypersensitivity / heart field specification / lymphocyte adhesion to endothelial cell of high endothelial venule / heparan sulfate N-acetylglucosaminyltransferase activity / glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase activity / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase activity / smoothened signaling pathway involved in lung development / developmental growth involved in morphogenesis / sweat gland development / perichondral bone morphogenesis / mesenchymal cell differentiation involved in bone development / response to leukemia inhibitory factor / UDP-N-acetylglucosamine transferase complex / chondrocyte hypertrophy / embryonic skeletal joint development / hematopoietic stem cell migration to bone marrow / fluid transport / limb joint morphogenesis / tight junction organization / glucuronosyltransferase activity / Defective EXT2 causes exostoses 2 / Defective EXT1 causes exostoses 1, TRPS2 and CHDS / heparin proteoglycan biosynthetic process / stomach development / sebaceous gland development / response to heparin / glomerular basement membrane development / glycosaminoglycan biosynthetic process / HS-GAG biosynthesis / glandular epithelial cell differentiation / lymphocyte migration into lymphoid organs / sulfation / chondrocyte proliferation / dendritic cell migration / endochondral bone morphogenesis / dendrite self-avoidance / heparan sulfate proteoglycan biosynthetic process / endochondral bone growth / acetylglucosaminyltransferase activity / hematopoietic stem cell homeostasis / podocyte differentiation / sodium ion homeostasis / cartilage development involved in endochondral bone morphogenesis / basement membrane organization / vocalization behavior / endochondral ossification / olfactory bulb development / vacuole organization / cranial skeletal system development / leukocyte tethering or rolling / multicellular organismal-level water homeostasis / endoderm development / polysaccharide biosynthetic process / fear response / response to light intensity / optic nerve development / protein N-linked glycosylation / regulation of tumor necrosis factor-mediated signaling pathway / collagen fibril organization / neural crest cell differentiation / ossification involved in bone maturation / stem cell division / hair follicle morphogenesis / cell adhesion mediated by integrin / protein glycosylation / heart contraction / epithelial tube branching involved in lung morphogenesis / antigen processing and presentation / glycosyltransferase activity / mesoderm development / motor behavior / mesoderm formation / blood vessel remodeling / social behavior / catalytic complex / cell fate commitment / fibroblast growth factor receptor signaling pathway / hematopoietic stem cell differentiation / chondrocyte differentiation / canonical Wnt signaling pathway / BMP signaling pathway / bone resorption / cellular response to fibroblast growth factor stimulus / gastrulation / ossification / axon guidance / synaptic transmission, glutamatergic / wound healing / protein catabolic process / cellular response to virus / regulation of blood pressure / multicellular organism growth / vasodilation / protein-containing complex assembly / gene expression / protein heterodimerization activity / Golgi membrane / synapse 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: 3.1 Å | ||||||
![]() | Li, H. / Li, H. | ||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structural basis for heparan sulfate co-polymerase action by the EXT1-2 complex. 著者: Hua Li / Digantkumar Chapla / Robert A Amos / Annapoorani Ramiah / Kelley W Moremen / Huilin Li / ![]() 要旨: Heparan sulfate (HS) proteoglycans are extended (-GlcAβ1,4GlcNAcα1,4-) co-polymers containing decorations of sulfation and epimerization that are linked to cell surface and extracellular matrix ...Heparan sulfate (HS) proteoglycans are extended (-GlcAβ1,4GlcNAcα1,4-) co-polymers containing decorations of sulfation and epimerization that are linked to cell surface and extracellular matrix proteins. In mammals, HS repeat units are extended by an obligate heterocomplex of two exostosin family members, EXT1 and EXT2, where each protein monomer contains distinct GT47 (GT-B fold) and GT64 (GT-A fold) glycosyltransferase domains. In this study, we generated human EXT1-EXT2 (EXT1-2) as a functional heterocomplex and determined its structure in the presence of bound donor and acceptor substrates. Structural data and enzyme activity of catalytic site mutants demonstrate that only two of the four glycosyltransferase domains are major contributors to co-polymer syntheses: the EXT1 GT-B fold β1,4GlcA transferase domain and the EXT2 GT-A fold α1,4GlcNAc transferase domain. The two catalytic sites are over 90 Å apart, indicating that HS is synthesized by a dissociative process that involves a single catalytic site on each monomer. | ||||||
履歴 |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
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PDBx/mmCIF形式 | ![]() | 232 KB | 表示 | ![]() |
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PDB形式 | ![]() | 177.5 KB | 表示 | ![]() |
PDBx/mmJSON形式 | ![]() | ツリー表示 | ![]() | |
その他 | ![]() |
-検証レポート
文書・要旨 | ![]() | 1.2 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.2 MB | 表示 | |
XML形式データ | ![]() | 45.9 KB | 表示 | |
CIF形式データ | ![]() | 69.6 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 25035MC ![]() 7scjC ![]() 7sckC ![]() 7uqxC ![]() 7uqyC M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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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: 83404.023 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() 参照: UniProt: Q16394, glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase, N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase |
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#2: タンパク質 | 分子量: 77238.031 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() 参照: UniProt: Q93063, glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase, N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase |
#3: 多糖 | beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta- ...beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose |
研究の焦点であるリガンドがあるか | Y |
Has protein modification | Y |
-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: PARTICLE / 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: hEXT1/2 / タイプ: COMPLEX / Entity ID: #1-#2 / 由来: RECOMBINANT | |||||||||||||||
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由来(天然) | 生物種: ![]() | |||||||||||||||
由来(組換発現) | 生物種: Mammalia (両生類) | |||||||||||||||
緩衝液 | pH: 7 | |||||||||||||||
緩衝液成分 |
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試料 | 濃度: 0.3 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | |||||||||||||||
試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: Quantifoil R2/1 | |||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 95 % / 凍結前の試料温度: 299 K |
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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 / 倍率(公称値): 105000 X / Cs: 2.7 mm / C2レンズ絞り径: 70 µm / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER 最高温度: 193 K / 最低温度: 193 K / Residual tilt: 0.05 mradians |
撮影 | 平均露光時間: 1.5 sec. / 電子線照射量: 66.8 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: 2980 |
画像スキャン | 横: 5760 / 縦: 4092 |
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解析
EMソフトウェア |
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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: 5186047 | ||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||
3次元再構成 | 解像度: 3.1 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 120216 / 対称性のタイプ: POINT | ||||||||||||||||||||
原子モデル構築 | プロトコル: AB INITIO MODEL |