Antiribophorin antibodies inhibit the targeting to the ER membrane of ribosomes containing nascent secretory polypeptides.
Polyclonal antibodies directed against ribophorins I and II, two membrane glycoproteins characteristic of the rough endoplasmic reticulum, inhibit the cotranslational translocation of a secretory protein growth hormone into the lumen of dog pancreas or rat liver microsomes. As expected, site-specific antibodies to epitopes located within the cytoplasmic domain of ribophorin I, but not antibodies to epitopes in the luminal domain of this protein, were effective in inhibiting translocation. Since monovalent Fab fragments were as inhibitory as intact IgG molecules, ribophorins must be closely associated with the translocation site and, therefore, are likely to function at some stage in the translocation process. In all cases, the antibodies that inhibited translocation also caused a significant reduction in total protein synthesis and treatments that neutralized their capacity to inhibit translocation also prevented their inhibitory effect on protein synthesis. This would be expected if the antibodies blocked the membrane-mediated relief of the SRP-induced arrest of polypeptide elongation. The antibodies were effective only when added before translocation was allowed to begin. In this case, they prevented the targeting of active ribosomes containing mRNA and nascent chains to the ER membrane. Thus, ribophorins must either directly participate in targeting or be so close to the targeting site that the antibodies sterically blocked this early phase of the translocation process.
Department of Cell Biology, New York University School of Medicine, New York 10016.
Y H
YH
Yu
D D
DD
Sabatini
G
G
Kreibich
D000818:Animals; D000906:Antibodies; D001692:Biological Transport / Q000502:physiology; D002474:Cell-Free System; D003593:Cytoplasm / Q000378:metabolism; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D013006:Growth Hormone / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments; D008565:Membrane Proteins / Q000502:physiology*; D008861:Microsomes / Q000378:metabolism; D010179:Pancreas / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000502:physiology*; D012270:Ribosomes / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013997:Time Factors
D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D008565:Membrane Proteins; D010455:Peptides; C034241:ribophorin; D013006:Growth Hormone