Interaction Of gD Research Articles

Cell Fusion Induced by Herpes Simplex Virus Glycoproteins gB, gD, and gH-gL Requires a gD Receptor but Not Necessarily Heparan Sulfate

To characterize cellular factors required for herpes simplex virus type 1 (HSV-1)-induced cell fusion, we used an efficient and quantitative assay relying on expression of HSV-1 glycoproteins in transfected cells. We showed the following: (1) Cell fusion depended not only on expression of four viral glycoproteins (gB, gD, and gH-gL), as previously shown, but also on expression of cell surface entry receptors specific for gD. (2) Cell fusion required expression of all four glycoproteins in the same cell. (3) Heparan sulfate was not required for cell fusion. (4) Coexpression of receptor with the four glycoproteins in the same cell reduced fusion activity, indicating that interaction of gD and receptor can limit polykaryocyte formation. Overall, the viral and cellular determinants of HSV-1-induced cell fusion are similar to those for viral entry, except that HSV-1 entry is significantly enhanced by binding of virus to cell surface heparan sulfate.

Interaction of Gd(III) MRI contrast agents with membranes: a review of recent EPR studies

Interaction of Gd(III) MRI contrast agents with membranes: a review of recent EPR studies

Interaction of Gd(III) MRI contrast agents with membranes: a review of recent EPR studies.

Rational development of new selective paramagnetic contrast agents (PCAs) requires a detailed understanding of their interactions with biological macromolecules. This report shows how some of these interactions can be studied with electron paramagnetic resonance (EPR) through examples of Gd3+ complexes interactions with model phospholipid membranes. It is shown that the spin label EPR method can be used to detect: (i) presence and possible location of lipophilic contrast agents in the model membranes, (ii) changes and distortions in membrane organization upon interaction with the PCAs, and (iii) changes in the local polarity of the bilayer and its phase behavior due to addition of Gd3+ complexes. This work demonstrates that interaction of Gd3+ complexes with phospholipid bilayers can be observed directly from changes in their continuous wave (CW) EPR spectra obtained at frequencies higher than X-band (9.5 GHz), where signals arising from aqueous and lipid-bound Gd3+ complexes become resolved. Analysis of frequency dependence of the effective g-factors of the EPR signal provides estimates of zero-field splitting (ZFS) parameter for these complexes at physiological conditions and information on how this parameter is affected by interaction with lipids. Multifrequency EPR experiments at high magnetic fields are also useful in providing data on the frequency dispersion of electronic relaxation caused mainly by a modulation of the electron-electron dipolar interaction (ZFS) of these high spin ions.

Anti-idiotypic antibodies mimicking glycoprotein D of herpes simplex virus identify a cellular protein required for virus spread from cell to cell and virus-induced polykaryocytosis.

Glycoprotein D (gD) of herpes simplex virus 1 (HSV-1) is required for stable attachment and penetration of the virus into susceptible cells after initial binding. We derived anti-idiotypic antibodies to the neutralizing monoclonal antibody HD1 to gD of HSV-1. These antibodies have the properties expected of antibodies against a gD receptor. Specifically, they bind to the surface of HEp-2, Vero, and HeLa cells susceptible to HSV infection and specifically react with a Mr 62,000 protein in these and other (143TK- and BHK) cell lines. They neutralize virion infectivity, drastically decrease plaque formation by impairing cell-to-cell spread of virions, and reduce polykaryocytosis induced by strain HFEM, which carries a syncytial (syn-) mutation. They do not affect HSV growth in a single-step cycle and plaque formation by an unrelated virus, indicating that they specifically affect the interaction of HSV gD) with a cell surface receptor. We conclude that the Mr 62,000 cell surface protein interacts with gD to enable spread of HSV-1 from cell to cell and virus-induced polykaryocytosis.

Role of glycoprotein gD in the adhesion of pseudorabies virus infected cells and subsequent cell-associated virus spread.

Pseudorabies virus (PrV) infected cells in suspension are able to adhere to a monolayer of uninfected cells by means of PrV glycoproteins expressed at the outer cell membrane, with gB and gC playing a major role as ligands and a heparinlike substance as receptor. In order to investigate the role of gD in this process and subsequent transmission of infectivity to contact cells, experiments with a gD deletion mutant, heparin and a monoclonal antibody (Mab) against gD were performed. The first indication that gD is active during cell adhesion was found by the observation that the binding of gD- PrV infected cells was five times weaker than that of wild type (WT) PrV infected cells. Further evidence was given by the use of a Mab against gD. Preincubation of WT PrV infected cells with this Mab led to a reduction of the percentage adhering cells from 69% to 49%. The same Mab inhibited the heparin independent and heparin resistant binding of WT PrV infected cells indicating that gD is important during both processes. Furthermore, it was demonstrated in a plaque assay that, after contact with a monolayer, gD- PrV infected cells in suspension were able to induce plaques with an efficiency of 1%. In conclusion, we can state that beside the interaction of the ligands gB and gC with a heparinlike receptor also the interaction of gD with a receptor which differs from a heparinlike substance mediates the binding of WT PrV infected cells to uninfected cells and that gD is not essential for the subsequent cell-to-cell spread of the virus.

Interaction of herpes simplex virus glycoprotein gC with mammalian cell surface molecules.

The entry of herpes simplex virus (HSV) into mammalian cells is a multistep process beginning with an attachment step involving glycoproteins gC and gB. A second step requires the interaction of glycoprotein gD with a cell surface molecule. We explored the interaction between gC and the cell surface by using purified proteins in the absence of detergent. Truncated forms of gC and gD, gC1(457t), gC2(426t), and gD1(306t), lacking the transmembrane and carboxyl regions were expressed in the baculovirus system. We studied the ability of these proteins to bind to mammalian cells, to bind to immobilized heparin, to block HSV type 1 (HSV-1) attachment to cells, and to inhibit plaque formation by HSV-1. Each of these gC proteins bound to conformation-dependent monoclonal antibodies and to human complement component C3b, indicating that they maintained the same conformation of gC proteins expressed in mammalian cells. Biotinylated gC1(457t) and gC2(426t) each bind to several cell lines. Binding was inhibited by an excess of unlabeled gC but not by gD, indicating specificity. The attachment of gC to cells involves primarily heparan sulfate proteoglycans, since heparitinase treatment of cells reduced gC binding by 50% but had no effect on gD binding. Moreover, binding of gC to two heparan sulfate-deficient L-cell lines, gro2C and sog9, both of which are mostly resistant to HSV infection, was markedly reduced. Purified gD1 (306t), however, bound equally well to the two mutant cell lines. In contrast, saturating amounts of gC1(457t) interfered with HSV-1 attachment to cells but failed to block plaque formation, suggesting a role for gC in attachment but not penetration. A mutant form of gC lacking residues 33 to 123, gC1(delta 33-123t), expressed in the baculovirus system, bound significantly less well to cells than did gC1(457t) and competed poorly with biotinylated gC1(457t) for binding. These results suggest that residues 33 to 123 are important for gC attachment to cells. In contrast, both the mutant and wild-type forms of gC bound to immobilized heparin, indicating that binding of these proteins to the cell surface involves more than a simple interaction with heparin. To determine that the contribution of the N-terminal region of gC is important for HSV attachment, we compared several properties of a mutant HSV-1 which contains gC lacking amino acids 33 to 123 to those of its parental virus, which contains full-length gC. The mutant bound less well to cells than the parental virus but exhibited normal growth properties.(ABSTRACT TRUNCATED AT 400 WORDS)

Soluble glycoprotein D blocks herpes simplex virus type 1 infection of rat eyes.

Herpes simplex virus type 1 (HSV-1) ocular infection in rats was blocked by treating the eyes with UV-inactivated virions containing glycoprotein D (gD) prior to ocular challenge. In contrast, rats treated with UV-inactivated virions lacking gD were not protected. A soluble, truncated form of HSV-2 gD (gD-2t) also protected against ocular infection. Treatment with gD-2t not only reduced mortality but also restricted progression of pathology and reduced the amount of viral antigen in the cornea. Host antibody or alpha/beta interferon responses to the gD-2t treatment were not detected. These results are similar to those observed in cell culture (D. C. Johnson, R. L. Burke, and T. Gregory, J. Virol. 64:2569-2576, 1990). The in vivo effect of exogenous gD is consistent with blocking of a cell surface gD receptor or with an inhibitory interaction of gD with virions.

Identification of herpes simplex virus type 1 glycoproteins interacting with the cell surface

To investigate the interaction of herpes simplex virus type 1 (HSV-1) with the cell surface, we studied the formation of complexes by HSV-1 virion proteins with biotinylated cell membrane components. HSV-1 virion proteins reactive with surface components of HEp-2 and other cells were identified as gC, gB, and gD. Results from competition experiments suggested that binding of gC, gB, and gD occurred in a noncooperative way. The observed complex formation could be specifically blocked by monospecific rabbit antisera against gB and gD. The interaction of gD with the cell surface was also inhibited by monoclonal antibody IV3.4., whereas other gD-specific monoclonal antibodies, despite their high neutralizing activity, were not able to inhibit this interaction. Taken together, these data provide direct evidence that at least three of the seven known HSV-1 glycoproteins are able to form complexes with cellular surface structures.

Interaction of rare earth metal ions with nucleosides—a study of binary and ternary systems in solution

Studies on interaction of La(III), Pr(III), Nd(III), Sm(III), Gd(III), Dy(III) and Er(III) with inosine and xanthosine in a 1:1 ratio have been carried out by potentiometric equilibrium measurements at 35±0·1°C and 0·1 M (KNO3) ionic strength. Investigations were also made for the interaction of these metal ions and nucleosides with the biologically important secondary ligands glycine and histidine. These investigations were undertaken with a view to assess the influence of charge on the structure and stability of 1:1 metal-inosine/xanthosine systems.

13C-n.m.r.-spectral study of the mode of interaction of Gd 3+ and Mn 2+ with two vicinally di- O- d-galactosylated tripeptides

Natural-abundance, 13C-n.m.r. spectroscopy was used to study the binding of Gd 3+ and Mn 2+ to two vicinally di- O-α- and -β- d-galactosylated tripeptides composed of Gly and l-Thr. Gd 3+ and Mn 2+ appear to interact with the α- d-Gal groups of the di- O-α- d-galactosylated tripeptide at two sites: near O-6′, and in the vicinity of O-2′ and Thr O-3. The metal-ion-binding to the β- d-Gal groups of the di- O-β- d-galactosylated tripeptide indicates that a strong binding-site exists near O-6′ and, possibly, several weak ones near O-3′ and Thr O-3. In the case of the di- O-α- d-galactosylated tripeptides, vicinal glycosylation appears to have little effect on the metal-ion-binding of the α- d-Gal groups.

13C-n.m.r.-spectral study of the mode of binding of Gd 3+ to various glycopeptides

Natural-abundance, 13C-n.m.r. spectroscopy was used to study the mode of binding of Gd 3+ to mono- O-glycosylated l-serine and tripeptides variously composed of Gly and l-Thr. When the amino and carboxyl groups of the amino acid are not blocked, strong interaction of Gd 3+ with them is observed; this is also readily apparent with some related, nonglycosylated peptides. When the amino and carboxyl groups of the amino acid are blocked, noticeable interaction of Gd 3+ with the glycosidic oxygen atom (O-3) and O-2′ for the glycopeptide containing α- d-Gal p, and with O-3 and N-2′ for the glycopeptide containing α- d-Gal pNAc, is observed. Weak interactions are also possible with O-4′ and O-6′ of the glycosyl groups. Although the amino acids were protected, these metal ion-carbohydrate interactions may still be mediated, to some extent, by the acetyl protecting the amino group and by the ester group on the amino acid.