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PMID: 2172972 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The overexpressed human 46-kDa mannose 6-phosphate receptor mediates endocytosis and sorting of beta-glucuronidase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 20 ·1990-10-00 ·Pages 8036-40

Watanabe H, Grubb JH, Sly WS

Abstract

We studied the function of the human small (46-kDa) mannose 6-phosphate receptor (SMPR) in transfected mouse L cells that do not express the larger insulin-like growth factor II/mannose 6-phosphate receptor. Cells overexpressing human SMPR were studied for enzyme binding to cell surface receptors, for binding to intracellular receptors in permeabilized cells, and for receptor-mediated endocytosis of recombinant human beta-glucuronidase. Specific binding to human SMPR in permeabilized cells showed a pH optimum between pH 6.0 and pH 6.5. Binding was significant in the presence of EDTA but was enhanced by added divalent cations. Up to 2.3% of the total functional receptor could be detected on the cell surface by enzyme binding. We present experiments showing that at very high levels of overexpression, and at pH 6.5, human SMPR mediated the endocytosis of beta-glucuronidase. At pH 7.5, the rate of endocytosis was only 14% the rate seen at pH 6.5. Cells overexpressing human SMPR also showed reduced secretion of newly synthesized beta-glucuronidase when compared to cells transfected with vector only, suggesting that overexpressed human SMPR can participate in sorting of newly synthesized beta-glucuronidase and partially correct the sorting defect in mouse L cells that do not express the insulin-like growth factor II/mannose 6-phosphate receptor.

MeSH Terms
Animals Cations, Divalent Cell Membrane Permeability Chromatography, Affinity Endocytosis Glucuronidase/metabolism Humans Kinetics L Cells/metabolism Mannosephosphates/metabolism Mice Molecular Weight Protein Binding Receptor, IGF Type 2 Receptors, Cell Surface/genetics,isolation & purification,metabolism Recombinant Proteins/metabolism Transfection
Chemicals
Cations, Divalent Mannosephosphates Receptor, IGF Type 2 Receptors, Cell Surface Recombinant Proteins Glucuronidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Watanabe H
Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, MO 63104.
Grubb J H
Sly W S
References (33)
33 references, click to expand
  1. Processing of the phosphorylated recognition marker in lysosomal enzymes. Characterization and partial purification of a microsomal alpha-N-acetylglucosaminyl phosphodiesterase.
    J Biol Chem. 1981 Jun 10;256(11):5717-21 PMID: 6263889
  2. UDP-N-acetylglucosamine:glycoprotein N-acetylglucosamine-1-phosphotransferase. Proposed enzyme for the phosphorylation of the high mannose oligosaccharide units of lysosomal enzymes.
    J Biol Chem. 1981 May 10;256(9):4275-81 PMID: 6452459
  3. 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
  4. Lysosomal enzyme targeting. N-Acetylglucosaminylphosphotransferase selectively phosphorylates native lysosomal enzymes.
    J Biol Chem. 1981 Dec 10;256(23):11977-80 PMID: 6457829
  5. UDP-N-acetylglucosamine:lysosomal enzyme precursor N-acetylglucosamine-1-phosphotransferase. Partial purification and characterization of the rat liver Golgi enzyme.
    J Biol Chem. 1982 Oct 25;257(20):12322-31 PMID: 6288715
  6. Identification and characterization of cells deficient in the mannose 6-phosphate receptor: evidence for an alternate pathway for lysosomal enzyme targeting.
    Proc Natl Acad Sci U S A. 1983 Feb;80(3):775-9 PMID: 6298775
  7. Studies of the biosynthesis of the mannose 6-phosphate receptor in receptor-positive and -deficient cell lines.
    J Cell Biol. 1983 Dec;97(6):1700-6 PMID: 6315741
  8. Antibody to mannose 6-phosphate specific receptor induces receptor deficiency in human fibroblasts.
    EMBO J. 1984 Jun;3(6):1281-6 PMID: 6086314
  9. 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
  10. Internalization of blocking antibodies against mannose-6-phosphate specific receptors.
    EMBO J. 1985 Jul;4(7):1725-30 PMID: 3161725
  11. 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
  12. 46 kd mannose 6-phosphate receptor: cloning, expression, and homology to the 215 kd mannose 6-phosphate receptor.
    Cell. 1987 Jul 17;50(2):181-92 PMID: 2954652
  13. Cloning of a cDNA encoding the human cation-dependent mannose 6-phosphate-specific receptor.
    Proc Natl Acad Sci U S A. 1987 Aug;84(16):5575-9 PMID: 2441386
  14. Insulin-like growth factor II receptor as a multifunctional binding protein.
    Nature. 1987 Sep 24-30;329(6137):301-7 PMID: 2957598
  15. 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
  16. Mr 46,000 mannose 6-phosphate specific receptor: its role in targeting of lysosomal enzymes.
    EMBO J. 1987 Sep;6(9):2677-81 PMID: 2960521
  17. The human cation-independent mannose 6-phosphate receptor. Cloning and sequence of the full-length cDNA and expression of functional receptor in COS cells.
    J Biol Chem. 1988 Feb 15;263(5):2553-62 PMID: 2963003
  18. Cloning and sequence analysis of the cation-independent mannose 6-phosphate receptor.
    J Biol Chem. 1988 Feb 15;263(5):2563-70 PMID: 2963004
  19. Antibody to the phosphomannosyl receptor inhibits recycling of receptor in fibroblasts.
    J Cell Biochem. 1987 Oct;35(2):137-51 PMID: 2828384
  20. The cation-independent mannose 6-phosphate receptor binds insulin-like growth factor II.
    J Biol Chem. 1988 Feb 25;263(6):2585-8 PMID: 2963812
  21. Proliferin secreted by cultured cells binds to mannose 6-phosphate receptors.
    J Biol Chem. 1988 Mar 5;263(7):3521-7 PMID: 2963825
  22. A single receptor binds both insulin-like growth factor II and mannose-6-phosphate.
    Science. 1988 Mar 4;239(4844):1134-7 PMID: 2964083
  23. The 'cation-dependent' mannose 6-phosphate receptor binds ligands in the absence of divalent cations.
    FEBS Lett. 1988 Sep 12;237(1-2):81-4 PMID: 2971570
  24. Expression of human cation-independent mannose 6-phosphate receptor cDNA in receptor-negative mouse P388D1 cells following gene transfer.
    J Biol Chem. 1988 Nov 5;263(31):16230-5 PMID: 2972705
  25. Mutations in the cytoplasmic domain of the 275 kd mannose 6-phosphate receptor differentially alter lysosomal enzyme sorting and endocytosis.
    Cell. 1989 Jun 2;57(5):787-96 PMID: 2541923
  26. Ligand interactions of the cation-dependent mannose 6-phosphate receptor. Comparison with the cation-independent mannose 6-phosphate receptor.
    J Biol Chem. 1989 May 15;264(14):7970-5 PMID: 2542255
  27. Quaternary structure of the Mr 46,000 mannose 6-phosphate specific receptor: effect of ligand, pH, and receptor concentration on the equilibrium between dimeric and tetrameric receptor forms.
    Biochemistry. 1990 Mar 13;29(10):2449-55 PMID: 2159321
  28. Binding of insulin-like growth factor II (IGF-II) by human cation-independent mannose 6-phosphate receptor/IGF-II receptor expressed in receptor-deficient mouse L cells.
    Cell Regul. 1990 Jan;1(2):197-213 PMID: 1966041
  29. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  30. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  31. Phosphohexosyl components of a lysosomal enzyme are recognized by pinocytosis receptors on human fibroblasts.
    Proc Natl Acad Sci U S A. 1977 May;74(5):2026-30 PMID: 266721
  32. Beta-glucuronidase binding to human fibroblast membrane receptors.
    J Biol Chem. 1980 Jun 10;255(11):5069-74 PMID: 6768729
  33. Purification and characterization of rat liver alpha-N-acetylglucosaminyl phosphodiesterase.
    J Biol Chem. 1981 Oct 10;256(19):9937-43 PMID: 6268636
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.20.8036
Published
1990-10-00
Pages
8036-40
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54887
Subset
IM
Grants
NIDDK NIH HHS · DK40163 · United States
NIGMS NIH HHS · GM34182 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.20.8036 PMC Full Text View journal details
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