ジャーナル: Elife / 年: 2020 タイトル: Tailored design of protein nanoparticle scaffolds for multivalent presentation of viral glycoprotein antigens. 著者: George Ueda / Aleksandar Antanasijevic / Jorge A Fallas / William Sheffler / Jeffrey Copps / Daniel Ellis / Geoffrey B Hutchinson / Adam Moyer / Anila Yasmeen / Yaroslav Tsybovsky / Young-Jun ...著者: George Ueda / Aleksandar Antanasijevic / Jorge A Fallas / William Sheffler / Jeffrey Copps / Daniel Ellis / Geoffrey B Hutchinson / Adam Moyer / Anila Yasmeen / Yaroslav Tsybovsky / Young-Jun Park / Matthew J Bick / Banumathi Sankaran / Rebecca A Gillespie / Philip Jm Brouwer / Peter H Zwart / David Veesler / Masaru Kanekiyo / Barney S Graham / Rogier W Sanders / John P Moore / Per Johan Klasse / Andrew B Ward / Neil P King / David Baker / 要旨: Multivalent presentation of viral glycoproteins can substantially increase the elicitation of antigen-specific antibodies. To enable a new generation of anti-viral vaccines, we designed self- ...Multivalent presentation of viral glycoproteins can substantially increase the elicitation of antigen-specific antibodies. To enable a new generation of anti-viral vaccines, we designed self-assembling protein nanoparticles with geometries tailored to present the ectodomains of influenza, HIV, and RSV viral glycoprotein trimers. We first designed trimers tailored for antigen fusion, featuring N-terminal helices positioned to match the C termini of the viral glycoproteins. Trimers that experimentally adopted their designed configurations were incorporated as components of tetrahedral, octahedral, and icosahedral nanoparticles, which were characterized by cryo-electron microscopy and assessed for their ability to present viral glycoproteins. Electron microscopy and antibody binding experiments demonstrated that the designed nanoparticles presented antigenically intact prefusion HIV-1 Env, influenza hemagglutinin, and RSV F trimers in the predicted geometries. This work demonstrates that antigen-displaying protein nanoparticles can be designed from scratch, and provides a systematic way to investigate the influence of antigen presentation geometry on the immune response to vaccination.
ダウンロード / ファイル: emd_21166.map.gz / 形式: CCP4 / 大きさ: 15.6 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
De novo designed icosahedral nanoparticle I53_dn5, Negative stain EM map
ボクセルのサイズ
X=Y=Z: 4.1 Å
密度
表面レベル
登録者による: 0.03 / ムービー #1: 0.03
最小 - 最大
-0.06897879 - 0.0835278
平均 (標準偏差)
-0.00020760664 (±0.0060626483)
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
160
160
160
Spacing
160
160
160
セル
A=B=C: 656.0 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
4.1
4.1
4.1
M x/y/z
160
160
160
origin x/y/z
0.000
0.000
0.000
length x/y/z
656.000
656.000
656.000
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-205
-205
-205
NX/NY/NZ
411
411
411
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
160
160
160
D min/max/mean
-0.069
0.084
-0.000
-
添付データ
-
試料の構成要素
-
全体 : De novo designed icosahedral nanoparticle I53_dn5, Negative Stain...
全体
名称: De novo designed icosahedral nanoparticle I53_dn5, Negative Stain EM Map
要素
複合体: De novo designed icosahedral nanoparticle I53_dn5, Negative Stain EM Map
タンパク質・ペプチド: I53_dn5A
タンパク質・ペプチド: I53_dn5B
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超分子 #1: De novo designed icosahedral nanoparticle I53_dn5, Negative Stain...
超分子
名称: De novo designed icosahedral nanoparticle I53_dn5, Negative Stain EM Map タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all 詳細: Nanoparticles were generated by co-expression of the two components (A and B) in E coli. Assembled particles were purified using a combination of Ni-affinity chromatography and gel filtration chromatography.
タイプ: NEGATIVE / 材質: Uranyl Formate 詳細: Sample diluted to 0.05 mg/mL. 3 uL was applied onto the grid, blotted off, and then stained with 2% uranyl formate for 60 seconds.