Abstract
Antibodies that block the ligand binding site of the cation-dependent mannose 6-phosphate specific receptor (Mr 46,000 MPR) were used to probe the function of the receptor in transport of lysosomal enzymes. Addition of the antibodies to the medium of Morris hepatoma 7777 cells, which express only the Mr 46,000 MPR, resulted in a decreased intracellular retention and increased secretion of newly synthesized lysosomal enzymes. In fibroblasts and HepG2 cells that express the cation-independent mannose 6-phosphate specific receptor (Mr 215,000 MPR) in addition to the Mr 46,000 MPR, antibodies against the Mr 46,000 MPR inhibited the intracellular retention of newly synthesized lysosomal enzymes only when added to the medium together with antibodies against the Mr 215,000 MPR. Morris hepatoma (M.H.) 7777 did not endocytose lysosomal enzymes, while U937 monocytes, which express both types of MPR, internalized lysosomal enzymes. The uptake was inhibited by antibodies against the Mr 215,000 MPR, but not by antibodies against the Mr 46,000 MPR. These observations suggest that Mr 46,000 MPR mediates transport of endogenous but not endocytosis of exogenous lysosomal enzymes. Internalization of receptor antibodies indicated that the failure to mediate endocytosis of lysosomal enzymes is due to an inability of surface Mr 46,000 MPR to bind ligands rather than its exclusion from the plasma membrane or from internalization.
MeSH Terms
Animals
Antibodies
Carrier Proteins/immunology,physiology
Cell Line
Endocytosis
Hexosephosphates/physiology
Humans
Kinetics
Liver Neoplasms, Experimental/enzymology
Lysosomes/enzymology
Mannosephosphates/physiology
Molecular Weight
Receptor, IGF Type 2
Chemicals
Antibodies
Carrier Proteins
Hexosephosphates
Mannosephosphates
Receptor, IGF Type 2
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Stein M
Physiologisch-Chemisches Institut, Universität Münster, FRG.
Zijderhand-Bleekemolen J E
Geuze H
Hasilik A
von Figura K
References (21)
21 references, click to expand
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
46-kDa mannose 6-phosphate-specific receptor: biosynthesis, processing, subcellular location and topology.
Biol Chem Hoppe Seyler. 1987 Aug;368(8):937-47
PMID: 2959299
-
A simplification of the protein assay method of Lowry et al. which is more generally applicable.
Anal Biochem. 1977 Dec;83(2):346-56
PMID: 603028
-
Biosynthesis of lysosomal enzymes in fibroblasts. Synthesis as precursors of higher molecular weight.
J Biol Chem. 1980 May 25;255(10):4937-45
PMID: 6989821
-
Characterization of a membrane-associated receptor from bovine liver that binds phosphomannosyl residues of bovine testicular beta-galactosidase.
Proc Natl Acad Sci U S A. 1981 Jul;78(7):4289-93
PMID: 6270668
-
Synthesis and processing of arylsulfatase A in human skin fibroblasts.
Hoppe Seylers Z Physiol Chem. 1982 Apr;363(4):425-30
PMID: 6122636
-
DIFFERENTIATION IN VITRO OF EMBRYONIC CARTILAGE AND BONE IN A CHEMICALLY-DEFINED MEDIUM.
Proc Soc Exp Biol Med. 1965 May;119:287-90
PMID: 14297872
-
Localization of the sites of iodination of human beta 2-microglobulin: quaternary structure implications for histocompatibility antigens.
Biochemistry. 1983 Mar 1;22(5):1145-53
PMID: 6188486
-
Biosynthesis and transport of cathepsin D in cultured human fibroblasts.
J Cell Biol. 1983 Jul;97(1):1-5
PMID: 6863385
-
Juvenile and adult metachromatic leukodystrophy: partial restoration of arylsulfatase A (cerebroside sulfatase) activity by inhibitors of thiol proteinases.
Proc Natl Acad Sci U S A. 1983 Oct;80(19):6066-70
PMID: 6136972
-
Antibody to mannose 6-phosphate specific receptor induces receptor deficiency in human fibroblasts.
EMBO J. 1984 Jun;3(6):1281-6
PMID: 6086314
-
Mannose 6-phosphate-specific receptor is a transmembrane protein with a C-terminal extension oriented towards the cytosol.
Biochem J. 1985 Jan 15;225(2):543-7
PMID: 3156583
-
Lysosomal enzyme binding to mouse P388D1 macrophage membranes lacking the 215-kDa mannose 6-phosphate receptor: evidence for the existence of a second mannose 6-phosphate receptor.
Proc Natl Acad Sci U S A. 1985 Jul;82(13):4428-32
PMID: 3160044
-
Transmembrane orientation of the mannose 6-phosphate receptor in isolated clathrin-coated vesicles.
J Biol Chem. 1985 Aug 15;260(17):9838-42
PMID: 2862145
-
Internalization of blocking antibodies against mannose-6-phosphate specific receptors.
EMBO J. 1985 Jul;4(7):1725-30
PMID: 3161725
-
Purification and characterization of a cation-dependent mannose 6-phosphate receptor from murine P388D1 macrophages and bovine liver.
J Biol Chem. 1985 Oct 5;260(22):12008-14
PMID: 2931431
-
Synthesis, transport and processing of cathepsin C in Morris hepatoma 7777 cells and rat hepatocytes.
Eur J Biochem. 1985 Nov 15;153(1):211-6
PMID: 4065148
-
Trafficking of lysosomal enzymes in normal and disease states.
J Clin Invest. 1986 Jan;77(1):1-6
PMID: 3003148
-
Lysosomal enzymes and their receptors.
Annu Rev Biochem. 1986;55:167-93
PMID: 2943218
-
Is movement of mannose 6-phosphate-specific receptor triggered by binding of lysosomal enzymes?
J Cell Biol. 1987 Jun;104(6):1735-42
PMID: 2953746
-
Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography.
Eur J Biochem. 1975 Aug 15;56(2):335-41
PMID: 1175627