Endosome acidification and receptor trafficking: bafilomycin A1 slows receptor externalization by a mechanism involving the receptor's internalization motif.
To examine the relationship between endosome acidification and receptor trafficking, transferrin receptor trafficking was characterized in Chinese hamster ovary cells in which endosome acidification was blocked by treatment with the specific inhibitor of the vacuolar H(+)-ATPase, bafilomycin A1. Elevating endosome pH slowed the receptor externalization rate to approximately one-half of control but did not affect receptor internalization kinetics. The slowed receptor externalization required the receptor's cytoplasmic domain and was largely eliminated by substitutions replacing either of two aromatic amino acids within the receptor's cytoplasmic YTRF internalization motif. These results confirm, using a specific inhibitor of the vacuolar proton pump, that proper endosome acidification is necessary to maintain rapid recycling of intracellular receptors back to the plasma membrane. Moreover, receptor return to the plasma membrane is slowed in the absence of proper endosome acidification by a signal-dependent mechanism involving the receptor's cytoplasmic tyrosine-containing internalization motif. These results, in conjunction with results from other studies, suggest that the mechanism for clustering receptors in plasma membrane clathrin-coated pits may be an example of a more general mechanism that determines the dynamic distribution of membrane proteins among various compartments with luminal acidification playing a crucial role in this process.
Johnson
L S
LS
Department of Pathology, Columbia University College of Physicians and Surgeons, New York, New York 10032.
Dunn
K W
KW
Pytowski
B
B
McGraw
T E
TE
D033942:Adaptor Proteins, Vesicular Transport; D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007424:Intracellular Fluid / Q000737:chemistry; D018942:Macrolides; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000378:metabolism; D015388:Organelles / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D017434:Protein Structure, Tertiary; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D011990:Receptors, Transferrin / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism
D033942:Adaptor Proteins, Vesicular Transport; D000900:Anti-Bacterial Agents; D018942:Macrolides; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D011990:Receptors, Transferrin; C057620:bafilomycin A; D006180:Proton-Translocating ATPases