National Institutes of Health/National Institute Of Allergy and Infectious Diseases
R01-AI104920
米国
引用
ジャーナル: Pathog Immun / 年: 2019 タイトル: α-Defensin HD5 Stabilizes Capsid/Core Interactions. 著者: Neetu M Gulati / Masaru Miyagi / Mayim E Wiens / Jason G Smith / Phoebe L Stewart / 要旨: BACKGROUND: (HPV) is linked to nearly all cases of cervical cancer. Despite available vaccines, a deeper understanding of the immune response to HPV is needed. Human α-defensin 5 (HD5), an innate ...BACKGROUND: (HPV) is linked to nearly all cases of cervical cancer. Despite available vaccines, a deeper understanding of the immune response to HPV is needed. Human α-defensin 5 (HD5), an innate immune effector peptide, blocks infection of multiple sero-types of HPV, including high-risk HPV16. While a common mechanism of α-defensin anti-viral activity against nonenveloped viruses such as HPV has emerged, there is limited understanding of how α-defensins bind to viral capsids to block infection. 手法: We have used cryo-electron microscopy (cryoEM), mass spectrometry (MS) crosslinking and differential lysine modification studies, and molecular dynamics (MD) simulations to probe the ...手法: We have used cryo-electron microscopy (cryoEM), mass spectrometry (MS) crosslinking and differential lysine modification studies, and molecular dynamics (MD) simulations to probe the interaction of HPV16 pseudovirions (PsVs) with HD5. RESULTS: CryoEM single particle reconstruction did not reveal HD5 density on the capsid surface. Rather, increased density was observed under the capsid shell in the presence of HD5. MS studies ...RESULTS: CryoEM single particle reconstruction did not reveal HD5 density on the capsid surface. Rather, increased density was observed under the capsid shell in the presence of HD5. MS studies indicate that HD5 binds near the L1 and L2 capsid proteins and specifically near the C-terminal region of L1. MD simulations indicate that favorable electrostatic interactions can be formed between HD5 and the L1 C-terminal tail. CONCLUSIONS: A model is presented for how HD5 affects HPV16 structure and cell entry. In this model, HD5 binds to disordered regions of L1 and L2 protruding from the icosahedrally ordered capsid. HD5 ...CONCLUSIONS: A model is presented for how HD5 affects HPV16 structure and cell entry. In this model, HD5 binds to disordered regions of L1 and L2 protruding from the icosahedrally ordered capsid. HD5 acts to cement interactions between L1 and L2 and leads to a closer association of the L2/genome core with the L1 capsid. This model provides a structural rationale for our prior observation that HD5 interferes with the separation of L1 from the L2/genome complex during cell entry.
ダウンロード / ファイル: emd_20218.map.gz / 形式: CCP4 / 大きさ: 824 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
CryoEM structure of human papillomavirus 16 pseudovirus in complex with human alpha-defensin 5 (HD5)
ボクセルのサイズ
X=Y=Z: 1.26 Å
密度
表面レベル
登録者による: 0.05 / ムービー #1: 0.08
最小 - 最大
-0.12733757 - 0.27827927
平均 (標準偏差)
0.0005827095 (±0.024572708)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
100
100
100
サイズ
600
600
600
Spacing
600
600
600
セル
A=B=C: 756.0 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
1.26
1.26
1.26
M x/y/z
600
600
600
origin x/y/z
0.000
0.000
0.000
length x/y/z
756.000
756.000
756.000
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
100
100
100
NC/NR/NS
600
600
600
D min/max/mean
-0.127
0.278
0.001
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添付データ
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試料の構成要素
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全体 : Complex of human papillomavirus type 16 pseudovirus with human al...
全体
名称: Complex of human papillomavirus type 16 pseudovirus with human alpha-defensin 5 (HD5)
要素
複合体: Complex of human papillomavirus type 16 pseudovirus with human alpha-defensin 5 (HD5)
複合体: alpha-defensin 5 (HD5)
ウイルス: Human papillomavirus type 16 (パピローマウイルス)
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超分子 #1: Complex of human papillomavirus type 16 pseudovirus with human al...
超分子
名称: Complex of human papillomavirus type 16 pseudovirus with human alpha-defensin 5 (HD5) タイプ: complex / ID: 1 / 親要素: 0
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超分子 #3: alpha-defensin 5 (HD5)
超分子
名称: alpha-defensin 5 (HD5) / タイプ: complex / ID: 3 / 親要素: 1 詳細: Synthesized linear HD5 peptide (CPC Scientific, Sunnyvale, CA) was subjected to thiol-disulfide reshuffling and purified to homogeneity by reverse-phase high-pressure liquid chromatography
由来(天然)
生物種: Homo sapiens (ヒト)
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超分子 #2: Human papillomavirus type 16
超分子
名称: Human papillomavirus type 16 / タイプ: virus / ID: 2 / 親要素: 1 / NCBI-ID: 333760 / 生物種: Human papillomavirus type 16 / Sci species strain: pseudovirus / ウイルスタイプ: VIRION / ウイルス・単離状態: OTHER / ウイルス・エンベロープ: No / ウイルス・中空状態: No
Host system
生物種: Homo sapiens (ヒト) / 組換細胞: 293TT
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実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
緩衝液
pH: 7.4
グリッド
詳細: unspecified
凍結
凍結剤: ETHANE
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: DIRECT ELECTRON DE-20 (5k x 3k) 平均電子線量: 60.0 e/Å2
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 FIELD
実験機器
モデル: Titan Krios / 画像提供: FEI Company
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画像解析
最終 再構成
解像度のタイプ: BY AUTHOR / 解像度: 4.9 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 1742