Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Oct 1;123(1):99–108. doi: 10.1083/jcb.123.1.99

Mannose 6-phosphate-independent targeting of lysosomal enzymes in I- cell disease B lymphoblasts

PMCID: PMC2119824  PMID: 8408210

Abstract

B lymphocytes from patients with I-cell disease (ICD) maintain normal cellular levels of lysosomal enzymes despite a deficiency of the enzyme UDP-N-acetylglucosamine: lysosomal enzyme N-acetylglucosamine-1- phosphotransferase. We find that an ICD B lymphoblastoid cell line targets about 45% of the lysosomal protease cathepsin D to dense lysosomes. This targeting occurs in the absence of detectable mannose 6- phosphate residues on the cathepsin D and is not observed in ICD fibroblasts. The secretory protein pepsinogen, which is closely related to cathepsin D in both amino acid sequence and three-dimensional structure, is mostly excluded from dense lysosomes, indicating that the lymphoblast targeting pathway is specific. Carbohydrate residues are not required for lysosomal targeting, since a non-glycosylated mutant cathepsin D is sorted with comparable efficiency to the wild type protein. Analysis of a number of cathepsin D/pepsinogen chimeric proteins indicates that an extensive polypeptide determinant in the cathepsin D carboxyl lobe can confer efficient lysosomal sorting when introduced into the pepsinogen sequence. This determinant overlaps but is not identical to the recognition marker for phosphotransferase. These results indicate that a specific protein recognition event underlies Man-6-P-independent lysosomal sorting in ICD lymphoblasts.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baranski T. J., Cantor A. B., Kornfeld S. Lysosomal enzyme phosphorylation. I. Protein recognition determinants in both lobes of procathepsin D mediate its interaction with UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase. J Biol Chem. 1992 Nov 15;267(32):23342–23348. [PubMed] [Google Scholar]
  2. Baranski T. J., Faust P. L., Kornfeld S. Generation of a lysosomal enzyme targeting signal in the secretory protein pepsinogen. Cell. 1990 Oct 19;63(2):281–291. doi: 10.1016/0092-8674(90)90161-7. [DOI] [PubMed] [Google Scholar]
  3. Baranski T. J., Koelsch G., Hartsuck J. A., Kornfeld S. Mapping and molecular modeling of a recognition domain for lysosomal enzyme targeting. J Biol Chem. 1991 Dec 5;266(34):23365–23372. [PubMed] [Google Scholar]
  4. Braulke T., Hasilik A., von Figura K. Low temperature blocks transport and sorting of cathepsin D in fibroblasts. Biol Chem Hoppe Seyler. 1988 Jun;369(6):441–449. doi: 10.1515/bchm3.1988.369.1.441. [DOI] [PubMed] [Google Scholar]
  5. Cantor A. B., Kornfeld S. Phosphorylation of Asn-linked oligosaccharides located at novel sites on the lysosomal enzyme cathepsin D. J Biol Chem. 1992 Nov 15;267(32):23357–23363. [PubMed] [Google Scholar]
  6. Davies D. R. The structure and function of the aspartic proteinases. Annu Rev Biophys Biophys Chem. 1990;19:189–215. doi: 10.1146/annurev.bb.19.060190.001201. [DOI] [PubMed] [Google Scholar]
  7. DiCioccio R. A., Miller A. L. Biosynthesis, processing, and secretion of alpha-L-fucosidase in lymphoid cells from patients with I-cell disease and pseudo-Hurler polydystrophy. Glycobiology. 1991 Dec;1(6):595–604. doi: 10.1093/glycob/1.6.595. [DOI] [PubMed] [Google Scholar]
  8. Diment S., Leech M. S., Stahl P. D. Cathepsin D is membrane-associated in macrophage endosomes. J Biol Chem. 1988 May 15;263(14):6901–6907. [PubMed] [Google Scholar]
  9. Faust P. L., Kornfeld S., Chirgwin J. M. Cloning and sequence analysis of cDNA for human cathepsin D. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4910–4914. doi: 10.1073/pnas.82.15.4910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Faust P. L., Wall D. A., Perara E., Lingappa V. R., Kornfeld S. Expression of human cathepsin D in Xenopus oocytes: phosphorylation and intracellular targeting. J Cell Biol. 1987 Nov;105(5):1937–1945. doi: 10.1083/jcb.105.5.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hasilik A., Neufeld E. F. Biosynthesis of lysosomal enzymes in fibroblasts. Phosphorylation of mannose residues. J Biol Chem. 1980 May 25;255(10):4946–4950. [PubMed] [Google Scholar]
  12. Hasilik A., Neufeld E. F. Biosynthesis of lysosomal enzymes in fibroblasts. Synthesis as precursors of higher molecular weight. J Biol Chem. 1980 May 25;255(10):4937–4945. [PubMed] [Google Scholar]
  13. Hasilik A., Waheed A., von Figura K. Enzymatic phosphorylation of lysosomal enzymes in the presence of UDP-N-acetylglucosamine. Absence of the activity in I-cell fibroblasts. Biochem Biophys Res Commun. 1981 Feb 12;98(3):761–767. doi: 10.1016/0006-291x(81)91177-3. [DOI] [PubMed] [Google Scholar]
  14. Hurtley S. M., Helenius A. Protein oligomerization in the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:277–307. doi: 10.1146/annurev.cb.05.110189.001425. [DOI] [PubMed] [Google Scholar]
  15. Johnson L. M., Bankaitis V. A., Emr S. D. Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease. Cell. 1987 Mar 13;48(5):875–885. doi: 10.1016/0092-8674(87)90084-5. [DOI] [PubMed] [Google Scholar]
  16. Klionsky D. J., Banta L. M., Emr S. D. Intracellular sorting and processing of a yeast vacuolar hydrolase: proteinase A propeptide contains vacuolar targeting information. Mol Cell Biol. 1988 May;8(5):2105–2116. doi: 10.1128/mcb.8.5.2105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kornfeld S., Mellman I. The biogenesis of lysosomes. Annu Rev Cell Biol. 1989;5:483–525. doi: 10.1146/annurev.cb.05.110189.002411. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Little L., Alcouloumre M., Drotar A. M., Herman S., Robertson R., Yeh R. Y., Miller A. L. Properties of N-acetylglucosamine 1-phosphotransferase from human lymphoblasts. Biochem J. 1987 Nov 15;248(1):151–159. doi: 10.1042/bj2480151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marciniszyn J., Jr, Huang J. S., Hartsuck J. A., Tang J. Mechanism of intramolecular activation of pepsinogen. Evidence for an intermediate delta and the involvement of the active site of pepsin in the intramolecular activation of pepsinogen. J Biol Chem. 1976 Nov 25;251(22):7095–7102. [PubMed] [Google Scholar]
  21. Margolskee R. F., Kavathas P., Berg P. Epstein-Barr virus shuttle vector for stable episomal replication of cDNA expression libraries in human cells. Mol Cell Biol. 1988 Jul;8(7):2837–2847. doi: 10.1128/mcb.8.7.2837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McIntyre G. F., Erickson A. H. Procathepsins L and D are membrane-bound in acidic microsomal vesicles. J Biol Chem. 1991 Aug 15;266(23):15438–15445. [PubMed] [Google Scholar]
  23. Miller S. G., Moore H. P. Regulated secretion. Curr Opin Cell Biol. 1990 Aug;2(4):642–647. doi: 10.1016/0955-0674(90)90105-n. [DOI] [PubMed] [Google Scholar]
  24. Okada S., Handa M., Hashimoto T., Nishimoto J., Inui K., Furukawa M., Furuyama J., Yabuuchi H., Tate M., Gasa S. Biochemical studies on lymphoblastoid cells with inherited N-acetyl-glucosamine 1-phosphotransferase deficiency (I-cell disease). Biochem Int. 1988 Aug;17(2):375–383. [PubMed] [Google Scholar]
  25. Owada M., Neufeld E. F. Is there a mechanism for introducing acid hydrolases into liver lysosomes that is independent of mannose 6-phosphate recognition? Evidence from I-cell disease. Biochem Biophys Res Commun. 1982 Apr 14;105(3):814–820. doi: 10.1016/0006-291x(82)91042-7. [DOI] [PubMed] [Google Scholar]
  26. Reitman M. L., Varki A., Kornfeld S. Fibroblasts from patients with I-cell disease and pseudo-Hurler polydystrophy are deficient in uridine 5'-diphosphate-N-acetylglucosamine: glycoprotein N-acetylglucosaminylphosphotransferase activity. J Clin Invest. 1981 May;67(5):1574–1579. doi: 10.1172/JCI110189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rijnboutt S., Kal A. J., Geuze H. J., Aerts H., Strous G. J. Mannose 6-phosphate-independent targeting of cathepsin D to lysosomes in HepG2 cells. J Biol Chem. 1991 Dec 15;266(35):23586–23592. [PubMed] [Google Scholar]
  28. Rijnboutt S., Stoorvogel W., Geuze H. J., Strous G. J. Identification of subcellular compartments involved in biosynthetic processing of cathepsin D. J Biol Chem. 1992 Aug 5;267(22):15665–15672. [PubMed] [Google Scholar]
  29. Schulze-Lohoff E., Hasilik A., von Figura K. Cathepsin D precursors in clathrin-coated organelles from human fibroblasts. J Cell Biol. 1985 Sep;101(3):824–829. doi: 10.1083/jcb.101.3.824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tsuji A., Omura K., Suzuki Y. I-cell disease: evidence for a mannose 6-phosphate independent pathway for translocation of lysosomal enzymes in lymphoblastoid cells. Clin Chim Acta. 1988 Aug 15;176(1):115–121. doi: 10.1016/0009-8981(88)90181-7. [DOI] [PubMed] [Google Scholar]
  31. Valls L. A., Winther J. R., Stevens T. H. Yeast carboxypeptidase Y vacuolar targeting signal is defined by four propeptide amino acids. J Cell Biol. 1990 Aug;111(2):361–368. doi: 10.1083/jcb.111.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Varki A., Kornfeld S. Structural studies of phosphorylated high mannose-type oligosaccharides. J Biol Chem. 1980 Nov 25;255(22):10847–10858. [PubMed] [Google Scholar]
  33. Varki A., Kornfeld S. The spectrum of anionic oligosaccharides released by endo-beta-N-acetylglucosaminidase H from glycoproteins. Structural studies and interactions with the phosphomannosyl receptor. J Biol Chem. 1983 Mar 10;258(5):2808–2818. [PubMed] [Google Scholar]
  34. Waheed A., Pohlmann R., Hasilik A., von Figura K., van Elsen A., Leroy J. G. Deficiency of UDP-N-acetylglucosamine:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase in organs of I-cell patients. Biochem Biophys Res Commun. 1982 Apr 14;105(3):1052–1058. doi: 10.1016/0006-291x(82)91076-2. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES