Internalization of blocking antibodies against mannose-6-phosphate specific receptors.
Antibodies against mannose-6-phosphate specific receptors inhibit the receptor-dependent endocytosis of exogenous lysosomal enzymes as well as the sorting of endogenous lysosomal enzymes. This inhibition was correlated with an apparent loss of the receptors. We report here that treatment of cells with the antibody results in the formation of receptor-antibody complexes that are not extracted by the procedure used for the solubilization of receptors prior to immunoprecipitation and detection of the receptor. The apparent loss of receptors is observed with both native antibody and the F(ab)2 fragments, but not with Fab fragments. In contrast the transport of lysosomal enzymes is inhibited by all three forms of the antibody. The inhibition is ascribed to masking by the antibody of the enzyme-binding site in the receptor. The inhibition of the sorting of endogenous lysosomal enzymes by antibodies added to the medium indicates that the mannose-6-phosphate specific receptors at the sorting site are in dynamic equilibrium with those at the cell surface. The receptor-antibody complexes formed at the cell surface appear to cycle between the cell surface and intracellular membranes. A fraction of the internalized antibodies dissociates from the receptors and is degraded after transfer into lysosomes. Complexing with Fab increases the concentration of the receptor in the lysosomes and decreases 2- to 3-fold the half-life of the receptor.
Gartung
C
C
Braulke
T
T
Hasilik
A
A
von Figura
K
K
D000906:Antibodies; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins / Q000276:immunology / Q000378:metabolism*; D004705:Endocytosis; D006600:Hexosephosphates / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008360:Mannosephosphates / Q000378:metabolism*; D017527:Receptor, IGF Type 2; D013347:Subcellular Fractions / Q000378:metabolism
D000906:Antibodies; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins; D006600:Hexosephosphates; D007140:Immunoglobulin Fab Fragments; D008360:Mannosephosphates; D017527:Receptor, IGF Type 2