ジャーナル: Nat Struct Mol Biol / 年: 2021 タイトル: Effect of SARS-CoV-2 B.1.1.7 mutations on spike protein structure and function. 著者: Tzu-Jing Yang / Pei-Yu Yu / Yuan-Chih Chang / Kang-Hao Liang / Hsian-Cheng Tso / Meng-Ru Ho / Wan-Yu Chen / Hsiu-Ting Lin / Han-Chung Wu / Shang-Te Danny Hsu / 要旨: The B.1.1.7 variant of SARS-CoV-2 first detected in the UK harbors amino-acid substitutions and deletions in the spike protein that potentially enhance host angiotensin conversion enzyme 2 (ACE2) ...The B.1.1.7 variant of SARS-CoV-2 first detected in the UK harbors amino-acid substitutions and deletions in the spike protein that potentially enhance host angiotensin conversion enzyme 2 (ACE2) receptor binding and viral immune evasion. Here we report cryo-EM structures of the spike protein of B.1.1.7 in the apo and ACE2-bound forms. The apo form showed one or two receptor-binding domains (RBDs) in the open conformation, without populating the fully closed state. All three RBDs were engaged in ACE2 binding. The B.1.1.7-specific A570D mutation introduces a molecular switch that could modulate the opening and closing of the RBD. The N501Y mutation introduces a π-π interaction that enhances RBD binding to ACE2 and abolishes binding of a potent neutralizing antibody (nAb). Cryo-EM also revealed how a cocktail of two nAbs simultaneously bind to all three RBDs, and demonstrated the potency of the nAb cocktail to neutralize different SARS-CoV-2 pseudovirus strains, including B.1.1.7.
The two deletions, including residues 69-70 and residue 144 are designed for this study. The ...The two deletions, including residues 69-70 and residue 144 are designed for this study. The residues, including Y498, D567, G611, H678, I713, A979 and H1115, are designed for this study. The residues 679-682 (RAAA) and residues 983-984 (PP) are designed for protein stabilization.
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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: SARS-CoV-2 spike glycoprotein / タイプ: ORGANELLE OR CELLULAR COMPONENT / 詳細: UK variant (B.1.1.7), two RBP-up conformation / Entity ID: #1 / 由来: RECOMBINANT
分子量
値: 0.54 MDa / 実験値: YES
由来(天然)
生物種: Severe acute respiratory syndrome coronavirus 2 (ウイルス)
由来(組換発現)
生物種: Homo sapiens (ヒト)
緩衝液
pH: 7.6
緩衝液成分
ID
濃度
名称
式
Buffer-ID
1
50mM
Tris
1
2
150mM
NaCl
1
3
0.02 %
sodiumazide
1
試料
濃度: 1 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES
急速凍結
装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K 詳細: blot for 2.5 seconds before plunging; blot force: 0; waiting time: 30s