Mutations in the cytoplasmic domain of the 275 kd mannose 6-phosphate receptor differentially alter lysosomal enzyme sorting and endocytosis.
The cation-independent mannose 6-phosphate receptor (Cl-MPR) sorts newly synthesized lysosomal enzymes in the Golgi and endocytoses extracellular lysosomal enzymes. To determine the role of the 163 amino acid cytoplasmic domain of the Cl-MPR in these functions, receptor-deficient mouse L cells were transfected with normal bovine Cl-MPR cDNA or cDNAs mutated in the cytoplasmic domain. The normal Cl-MPR functioned in sorting and endocytosis. Mutant receptors with 40 and 89 residues deleted from the carboxyl terminus of the cytoplasmic tail functioned normally in endocytosis, but were partially impaired in sorting. Mutant receptors with larger deletions leaving only 7 and 20 residues of the cytoplasmic tail were defective in endocytosis and sorting. A mutant receptor containing alanine instead of tyrosine residues at positions 24 and 26 was defective in endocytosis, and partially impaired in sorting. Receptors deficient in endocytosis accumulated at the cell surface. These results indicate that the cytoplasmic domain of the Cl-MPR contains different signals for rapid endocytosis and efficient lysosomal enzyme sorting.
Lobel
P
P
Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Fujimoto
K
K
Ye
R D
RD
Griffiths
G
G
Kornfeld
S
S
D000595:Amino Acid Sequence; D000818:Animals; D002402:Cathepsin D / Q000235:genetics; D003593:Cytoplasm / Q000378:metabolism; D004705:Endocytosis; D006600:Hexosephosphates / Q000378:metabolism*; D007739:L Cells / Q000201:enzymology / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology*; D008360:Mannosephosphates / Q000378:metabolism*; D051379:Mice; D008854:Microscopy, Electron; D008970:Molecular Weight; D009154:Mutation; D010957:Plasmids; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
D006600:Hexosephosphates; D008360:Mannosephosphates; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface; D001616:beta-Galactosidase; D002402:Cathepsin D