National Institutes of Health/National Center for Research Resources (NIH/NCRR)
R01GM102498
米国
National Institutes of Health/National Center for Research Resources (NIH/NCRR)
R01GM098672
米国
National Institutes of Health/National Center for Research Resources (NIH/NCRR)
S10OD020054
米国
National Institutes of Health/National Center for Research Resources (NIH/NCRR)
S10OD021741
米国
National Institutes of Health/National Center for Research Resources (NIH/NCRR)
R35GM122530
米国
引用
ジャーナル: Cell / 年: 2018 タイトル: Structural Basis for Cholesterol Transport-like Activity of the Hedgehog Receptor Patched. 著者: Yunxiao Zhang / David P Bulkley / Yao Xin / Kelsey J Roberts / Daniel E Asarnow / Ashutosh Sharma / Benjamin R Myers / Wonhwa Cho / Yifan Cheng / Philip A Beachy / 要旨: Hedgehog protein signals mediate tissue patterning and maintenance by binding to and inactivating their common receptor Patched, a 12-transmembrane protein that otherwise would suppress the activity ...Hedgehog protein signals mediate tissue patterning and maintenance by binding to and inactivating their common receptor Patched, a 12-transmembrane protein that otherwise would suppress the activity of the 7-transmembrane protein Smoothened. Loss of Patched function, the most common cause of basal cell carcinoma, permits unregulated activation of Smoothened and of the Hedgehog pathway. A cryo-EM structure of the Patched protein reveals striking transmembrane domain similarities to prokaryotic RND transporters. A central hydrophobic conduit with cholesterol-like contents courses through the extracellular domain and resembles that used by other RND proteins to transport substrates, suggesting Patched activity in cholesterol transport. Cholesterol activity in the inner leaflet of the plasma membrane is reduced by PTCH1 expression but rapidly restored by Hedgehog stimulation, suggesting that PTCH1 regulates Smoothened by controlling cholesterol availability.
THE AUTHORS STATE THAT THE EXACT IDENTITY OF THE LIGAND IS UNKNOWN. CHOLESTEROL WAS MODELED INTO ...THE AUTHORS STATE THAT THE EXACT IDENTITY OF THE LIGAND IS UNKNOWN. CHOLESTEROL WAS MODELED INTO THE CHOLESTEROL-LIKE DENSITY PRESENT IN THE MAP.
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実験情報
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実験
実験
手法: 電子顕微鏡法
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: Patched1 protein solubilized in amphipol / タイプ: COMPLEX / Entity ID: #1 / 由来: RECOMBINANT
分子量
値: 0.1452 MDa / 実験値: NO
由来(天然)
生物種: Homo sapiens (ヒト)
由来(組換発現)
生物種: Homo sapiens (ヒト) / 細胞: HEK293 / プラスミド: BacMam
緩衝液
pH: 7.4
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