Expression of antibody interferes with disulfide bond formation and intracellular transport of antigen in the secretory pathway. |
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Authors: | L Pelkmans A Helenius |
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Institution: | Laboratory for Biochemistry, Swiss Federal Institute of Technology Zürich, Universit?tstrasse 16, CH-8092 Zürich, Switzerland. |
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Abstract: | To determine whether antibodies would interfere with the folding of glycoprotein antigens in the endoplasmic reticulum lumen of living cells, hybridoma cells producing monoclonal anti-hemagglutinin (HA) antibodies were infected with influenza virus. The fate of the newly synthesized HA was determined using an established pulse-chase approach. When the monoclonal antibodies were against epitopes present on early folding intermediates, folding and intracellular transport of HA to the Golgi complex were severely disturbed. On the other hand, when the antibodies were specific for the native HA trimers, immune complexes were formed, but folding or transport of HA was not affected. The use of antibodies in this way provided in situ information about the protein folding process inside the endoplasmic reticulum lumen of cells without external perturbation of the folding chains or the folding compartment. |
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