ジャーナル: Sci Adv / 年: 2022 タイトル: Structural basis for HCMV Pentamer recognition by neuropilin 2 and neutralizing antibodies. 著者: Daniel Wrapp / Xiaohua Ye / Zhiqiang Ku / Hang Su / Harrison G Jones / Nianshuang Wang / Akaash K Mishra / Daniel C Freed / Fengsheng Li / Aimin Tang / Leike Li / Dabbu Kumar Jaijyan / Hua ...著者: Daniel Wrapp / Xiaohua Ye / Zhiqiang Ku / Hang Su / Harrison G Jones / Nianshuang Wang / Akaash K Mishra / Daniel C Freed / Fengsheng Li / Aimin Tang / Leike Li / Dabbu Kumar Jaijyan / Hua Zhu / Dai Wang / Tong-Ming Fu / Ningyan Zhang / Zhiqiang An / Jason S McLellan / 要旨: Human cytomegalovirus (HCMV) encodes multiple surface glycoprotein complexes to infect a variety of cell types. The HCMV Pentamer, composed of gH, gL, UL128, UL130, and UL131A, enhances entry into ...Human cytomegalovirus (HCMV) encodes multiple surface glycoprotein complexes to infect a variety of cell types. The HCMV Pentamer, composed of gH, gL, UL128, UL130, and UL131A, enhances entry into epithelial, endothelial, and myeloid cells by interacting with the cell surface receptor neuropilin 2 (NRP2). Despite the critical nature of this interaction, the molecular determinants that govern NRP2 recognition remain unclear. Here, we describe the cryo-EM structure of NRP2 bound to Pentamer. The high-affinity interaction between these proteins is calcium dependent and differs from the canonical carboxyl-terminal arginine (CendR) binding that NRP2 typically uses. We also determine the structures of four neutralizing human antibodies bound to the HCMV Pentamer to define susceptible epitopes. Two of these antibodies compete with NRP2 binding, but the two most potent antibodies recognize a previously unidentified epitope that does not overlap the NRP2-binding site. Collectively, these findings provide a structural basis for HCMV tropism and antibody-mediated neutralization.
The primary map is the result of a focused refinement that encompasses the HCMV UL proteins (UL128, UL130, UL131) and NRP2 domains a2b1b2. The included additional map is the non-focused refinement in which the entire HCMV pentamer ectodomain and the entirety of NRP2 a1a2b1b2 can be observed.
#1-#4
0
MULTIPLESOURCES
2
TheectodomainoftheHCMVpentamer
COMPLEX
#1-#3
1
RECOMBINANT
3
Domainsa1a2b1b2ofhumanneuropilin2
COMPLEX
#4
1
RECOMBINANT
分子量
値: 0.23 MDa / 実験値: YES
由来(天然)
ID
Entity assembly-ID
生物種
Ncbi tax-ID
2
2
Human betaherpesvirus 5 (ヘルペスウイルス)
10359
3
3
Homo sapiens (ヒト)
9606
由来(組換発現)
ID
Entity assembly-ID
生物種
Ncbi tax-ID
2
2
Homo sapiens (ヒト)
9606
3
3
Homo sapiens (ヒト)
9606
緩衝液
pH: 8
緩衝液成分
ID
濃度
名称
式
Buffer-ID
1
2mM
Tris
C4H11NO3
1
2
200mM
sodiumchloride
NaCl
1
3
2mM
calciumchloride
CaCl2
1
4
0.02 %
sodiumazide
NaN3
1
5
0.1 %
amphipol A8-35
(C6.2H10.3O1.35N0.65Na0.35)~72
1
試料
濃度: 0.4 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 / グリッドのタイプ: UltrAuFoil R1.2/1.3
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: 277.15 K / 詳細: Grid was blotted with a force of -6 for 3 seconds
-
電子顕微鏡撮影
実験機器
モデル: Titan Krios / 画像提供: FEI Company
顕微鏡
モデル: FEI TITAN KRIOS / 詳細: Collected from a single grid at -30 degrees tilt