Effect of anti-ER antibodies within the ER lumen of living cells.
We describe the production and partial characterization of 12 monoclonal antibodies raised against a preparation of endoplasmic reticulum membranes obtained from Xenopus laevis liver. Four of the antibodies cross-react with liver melanocytes; two of the antibodies recognize extracellular antigens, whilst the remaining six recognize antigens present in hepatocytes. The concentrations of these latter antigens increase markedly in livers stimulated by estrogen. Western blotting analysis revealed that the six anti-hepatocyte monoclonal antibodies recognize at least five different antigens whose molecular weights are 14K, 18K, 19K, 43K, and 125K. The possible functional involvement of the various antigens in the secretory pathway was investigated using Xenopus oocytes as a surrogate secretory system. The mRNAs coding for the monoclonal antibodies were injected into oocytes and the resulting immunoglobulin chains were translated and assembled into active anti-ER antibodies inside the lumen of the ER. The effect on secretion was then observed. Our data indicate that the binding of antibodies to most antigens of the endoplasmic reticulum membrane may result in a blockage of secretion.
Valle
G
G
Department of Biological Sciences, University of Warwick, Coventry, United Kingdom.
Bhamra
S S
SS
Martin
S
S
Griffiths
G
G
Colman
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D004958:Estradiol; D005260:Female; D006825:Hybridomas; D007150:Immunohistochemistry; D007158:Immunologic Techniques; D008099:Liver / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D008845:Microinjections; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D009113:Muramidase / Q000276:immunology / Q000378:metabolism; D009865:Oocytes / Q000276:immunology / Q000378:metabolism; D010047:Ovalbumin / Q000276:immunology / Q000378:metabolism; D012333:RNA, Messenger / Q000008:administration & dosage; D014819:Vitellogenins / Q000096:biosynthesis / Q000276:immunology; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D014819:Vitellogenins; D004958:Estradiol; D010047:Ovalbumin; D009113:Muramidase