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Biomedical subjects

A Scheid

Publications and source records attributed to A Scheid.

52 records · Page 3Linked to original sources

Role of paramyxovirus glycoproteins in the interactions between viral and cell membranes.

Interactions of paramyxoviruses with cell membranes are mediated by two virus-coded glycoproteins of the virus membrane: HN and F. The HN protein is responsible for the attachment of virions to the membrane of the target cell. The F protein mediates fusion between the virus membrane and the cell membrane, and this step is essential for the infection process as well as for the expression of other biological activities of paramyxoviruses, i.e., cell fusion and hemolysis. The activity of F is dependent on the proteolytic processing of a precursor (F0) to yield two disulfide-linked subunits (F1 and F2), and unless this cleavage occurs, the virions are inactive and not infectious. Viruses differ in their susceptibility to specific proteases, as demonstrated with Sendai virus and its protease activation (pa) mutants. Findings with these viruses indicate that the host range, tissue tropism, and the ability of paramyxoviruses to spread within a host and to cause disease are dependent on the availability of an appropriate protease capable of cleaving the viral glycoprotein. Experiments with reconstituted particles that contain purified F protein and phosphatidylcholine indicate that to be active the F protein must be inserted in a lipid bilayer, and that no other viral function is needed for the membrane fusion process. Attachment of the F-protein-containing particles to the target membrane is a prerequisite for fusion; however, this function can be supplied by wheat germ agglutinin as well as by the virus protein responsible for attachment of intact virus, i.e., HN. Studies on the structure of the paramyxovirus glycoproteins indicate that they are inserted with a hydrophobic region of the molecule in the lipid bilayer of the virus membrane. Analysis of the primary structure of the F protein at the cleavage site revealed striking sequence homology among three paramyxoviruses, indicating a requirement for cleavage at a sharply defined site and the importance of a specific primary sequence for biological activity. This region is highly hydrophobic, suggesting that it may interact with the lipid biylayer of the target cell membrane during virus penetration, cell fusion, or hemolysis. Further studies of this system should provide knowledge regarding the mechanisms involved in viral penetration and in membrane fusion in general.

Amino Acid Sequence↗

Loss on serial passage of rhesus monkey kidney cells of proteolytic activity required for Sendai virus activation.

Primary and secondary cultures of rhesus monkey kidney cells supported multiple-cycle replication of Sendai virus, but later passages lost this ability, and this was reflected in decreased plaque formation. Multiple-cycle replication also did not occur in LLC-MK2 cells, a continuous line of RMK cells. Failure of replication in serially passed cells was correlated with a decrease in proteolytic cleavage of a viral surface glycoprotein (Fo), and the ability of cells to support multiple-cycle replication and plaque formation could be restored by the addition of trypsin (0.3 microgram/ml) to the overlay medium. The use of wild-type virus, which requires trypsin, and protease activation mutants that require chymotrypsin or elastase for activation has provided evidence that the activating protease supplied by primary or secondary cells has trypsin-like activity. Inactive virus, with uncleaved Fo glycoprotein, absorbed to primary or secondary cells but did not infect them, even though such cells possess the enzyme that is capable of cleaving the Fo glycoprotein of virus synthesized in these cells. The inability of these cells to activate adsorbed virus indicates that the activating protease that they possess is inacessible to adsorbed virus, although it can act on the Fo glycoprotein during virus maturation in these cells. These data provide a biochemical explanation for the failure of later passages of a cell strain or a continuous cell line to support the replication of a paramyxovirus.

Animals↗

Isolation and purification of the envelope proteins of Newcastle disease virus.

A procedure has been developed for the isolation of Newcastle disease virus (NDV) envelope proteins. The two surface glycoproteins and the non-glycosylated membrane protein were solubilized with 2% Triton X-100 and 1 m KCl. Removal of the KCl by dialysis yielded by precipitation a pure preparation of the non-glycosylated membrane protein, which is insoluble in solutions of low ionic strength. The soluble fraction consisting of the two glycoproteins possessed full neuraminidase and hemagglutinating activities. The two glycoproteins could be separated by rate zonal sedimentation in a sucrose gradient containing 1% Triton X-100 and 1 m KCl. Under these conditions, the sedimentation coefficient of the larger glycoprotein, virus protein 1, was 9.3s, and that of the smaller, virus protein 2, was 6.1s. Both hemagglutinating and neuraminidase activities were associated with virus protein 1; virus protein 2 had neither activity. The results suggest that both activities reside on a single NDV glycoprotein. Similar results were obtained previously with another paramyxovirus, simian virus 5. These findings suggest that the association of hemagglutinating and neuraminidase activities with one glycoprotein is a general property of the paramyxovirus group.

Amino Acids↗

Circulatory changes after surfactant bolus instillation in lung-lavaged adult rabbits.

Following surfactant instillation in infants treated for respiratory distress syndrome, a mean arterial blood pressure (MABP) decrease is often observed. Its etiology and pathogenesis are still unknown. In this study various circulatory parameters were recorded continuously after surfactant instillation to elucidate the role of pulmonary vascular resistance as one possible cause for the MABP drop. Seven anesthetized adult New Zealand white rabbits were artificially ventilated after tracheotomy. Arterial and right atrial pressure were recorded continuously. Pulmonary artery pressure and cardiac output were determined by means of a thermodilution catheter. After inducing surfactant deficiency by repeated saline lavages, 200 mg/kg body weight of a natural surfactant preparation was administered by tracheal bolus instillation. PaO2 increased rapidly from 8.0 +/- 1.3 kPa to 51.2 +/- 8.8 kPa (mean +/- standard deviation) within 2 min (p < .05). MABP dropped from 12.1 +/- 1.9 kPa to 8.9 +/- 2.3 kPa within 2 min (p < .05). Pulmonary artery pressure, cardiac output, and right atrial pressure did not change during the observation period of 60 min. The results suggest that a peripheral vasodilatation is the most likely cause for the drop in MABP.

Animals↗