PDBsum entry 1jma

Viral protein

PDB id: 1jma

Contents

Protein chains

100 a.a. *

259 a.a. *

Ligands

NAG-NAG

SO4 ×5

Waters ×49

* Residue conservation analysis

References listed in PDB file

Key reference

Title

Herpes simplex virus glycoprotein d bound to the human receptor hvea.

Authors

A.Carfí, S.H.Willis, J.C.Whitbeck, C.Krummenacher, G.H.Cohen, R.J.Eisenberg, D.C.Wiley.

Ref.

Mol Cell, 2001, 8, 169-179. [DOI no: 10.1016/S1097-2765(01)00298-2]

PubMed id

11511370

Abstract

Herpes simplex virus (HSV) infection requires binding of the viral envelope glycoprotein D (gD) to cell surface receptors. We report the X-ray structures of a soluble, truncated ectodomain of gD both alone and in complex with the ectodomain of its cellular receptor HveA. Two bound anions suggest possible binding sites for another gD receptor, a 3-O-sulfonated heparan sulfate. Unexpectedly, the structures reveal a V-like immunoglobulin (Ig) fold at the core of gD that is closely related to cellular adhesion molecules and flanked by large N- and C-terminal extensions. The receptor binding segment of gD, an N-terminal hairpin, appears conformationally flexible, suggesting that a conformational change accompanying binding might be part of the viral entry mechanism.

Figure 3.
Figure 3. The gD-HveA Interface

Figure 5.
Figure 5. Conformational Change in the N Terminus of gD

The above figures are reprinted by permission from Cell Press: Mol Cell (2001, 8, 169-179) copyright 2001.

Secondary reference #1

Title

Herpes simplex virus-1 entry into cells mediated by a novel member of the tnf/ngf receptor family.

Authors

R.I.Montgomery, M.S.Warner, B.J.Lum, P.G.Spear.

Ref.

Cell, 1996, 87, 427-436. [DOI no: 10.1016/S0092-8674(00)81363-X]

PubMed id

8898196

Figure 3.
Figure 3. Southern and Northern Blots for Detection of HVEM–Homologous DNA and RNA in Cells and Tissues(A–C) Cell DNAs were extracted and digested with BamHI and the fragments separated by electrophoresis and transferred to Duralon nylon membrane for hybridization. The DNAs were from HeLa (lanes 2), HEp-2 (lanes 3), CHO-K1 (lanes 4), CHO-HVEM12 (lanes 5), and Vero (lanes 6) cells. BamHI-digested plasmid, pBEC580, was also included (lanes 7).(A) Photograph of the ethidium bromide–stained gel.(B) Autoradiogram of the blot using the PvuII probe indicated in (D).(C) Autoradiogram of the blot using the EcoRI probe indicated in (D).(D) Schematic diagram of the HVEM cDNA and fragments used to generate probes. Numbers on the left of (A) and bands in lane 1 indicate molecular size markers (kbp).(E) A Northern blot (Clontech) of polyadenylated RNAs extracted from various human tissues and hybridized with a ^32P-labeled probe from the PvuII fragment indicated in (D). The RNAs were extracted from heart (lane 1), brain (lane 2), placenta (lane 3), lung (lane 4), liver (lane 5), skeletal muscle (lane 6), kidney (lane 7), and pancreas (lane 8). Amounts on the blot were normalized with respect to actin mRNA content.

Figure 5.
Figure 5. Enhanced Entry of HSV-1(KOS) into HVEM–Expressing Cell Lines and Inhibition of Infection by Anti–HVEM Antibodies or HVEM:Fc(A) Several HVEM–expressing and control cell lines were obtained by transfection of CHO-K1 cells or ST cells with pBEC10 or control plasmid pcDNA3, followed by selection for stable maintenance of the plasmid. Representative clones were plated in 96 well plates and exposed to KOS-gL86 at the doses indicated. Later (6 hr), viral entry was quantitated as described in the legend to Figure 1. CHO-HVEM11 and ST-HVEM1 (open circles), CHO-HVEM9 and ST-HVEM22 (closed circles), and CHO-HVEM12 and ST-HVEM2 (open squares) were isolated after transfection with the HVEM–expressing plasmid pBEC10. CHO-C8 and ST-C8 (closed squares) were isolated after transfection with the control plasmid pcDNA3.(B and C) CHO-HVEM12 cells or ST-HVEM1 cells were plated in 96 well dishes. In (B), the cells were exposed to preimmune or immune rabbit serum at the dilutions indicated for 30 min at 37°C. Various amounts of KOS-gL86 were then added in one-fifth volume, and incubation continued for 2 hr. In (C), virus was mixed with various concentrations of HVEM:Fc or normal rabbit IgG and incubated for 30 min at 37°C. The mixtures (50 μl) were added to washed cells, and incubation continued for 2 hr. The virus–serum or virus–HVEM:Fc mixtures were then removed and the cells exposed briefly to low pH buffer to inactivate residual extracellular virus. The cells were washed and incubated for an additional 4 hr before lysis and addition of ONPG. The amount of virus added was 10^7 pfu/well in (B) and 10^6 pfu/well in (C). Open symbols were preimmune serum (B) or normal rabbit IgG (C). Closed symbols were immune serum (B) or HVEM:Fc (C).

The above figures are reproduced from the cited reference with permission from Cell Press