Multivesicular endosomes containing internalized EGF-EGF receptor complexes mature and then fuse directly with lysosomes.
We have followed the transfer of EGF-EGF receptor (EGFR) complexes from endosomal vacuoles that contain transferrin receptors (TfR) to lysosome vacuoles identified by their content of HRP loaded as a 15-min pulse 4 h previously. We show that the HRP-loaded lysosomes are lysosomal-associated membrane protein-1 (LAMP-1) positive, mannose-6-phosphate receptor (M6PR) negative. and contain active acid hydrolase. EGF-EGFR complexes are delivered to these lysosomes intact and are then rapidly degraded. Preactivating the HRP contained within the preloaded lysosomes inhibits the delivery of EGFR and degradation of EGF, and results in the accumulation of EGFR-containing multivesicular bodies (MVB). With time these accumulating MVB undergo a series of maturation changes that include the loss of TfR, the continued recruitment of EGFR, and the accumulation of internal vesicles, but they remain LAMP-1 and M6PR negative. The mature MVB are often seen to make direct contact with lysosomes containing preactivated HRP, but their perimeter membranes remain intact. Together our observations suggest that the transfer of EGF-EGFR complexes from the TfR-containing endosome compartment to the lysosomes that degrade them employs a single vacuolar intermediate, the maturing MVB, and can be achieved by a single heterotypic fusion step.
Futter
C E
CE
Medical Research Council Laboratory for Molecular Cell Biology, University College, London, United Kingdom.
Pearse
A
A
Hewlett
L J
LJ
Hopkins
C R
CR
D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers; D002460:Cell Line; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism* / Q000648:ultrastructure; D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D006868:Hydrolysis; D051907:Lysosomal Membrane Proteins; D008247:Lysosomes / Q000378:metabolism* / Q000648:ultrastructure; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000378:metabolism; D008854:Microscopy, Electron; D017527:Receptor, IGF Type 2 / Q000378:metabolism; D011990:Receptors, Transferrin / Q000032:analysis; D011994:Recombinant Proteins / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
D015703:Antigens, CD; D015415:Biomarkers; D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins; D017527:Receptor, IGF Type 2; D011990:Receptors, Transferrin; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D006735:Horseradish Peroxidase; D066246:ErbB Receptors