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. 1984 Nov;3(11):2671–2676. doi: 10.1002/j.1460-2075.1984.tb02192.x

Myeloperoxidase is synthesized as larger phosphorylated precursor.

A Hasilik, R Pohlmann, R L Olsen, K von Figura
PMCID: PMC557748  PMID: 6096138

Abstract

Synthesis and processing of myeloperoxidase were examined in metabolically labeled cells of the human promyelocyte line HL-60 and in an in vitro rabbit reticulocyte lysate system directed with HL-60 mRNA. Radioactivity labeled products were isolated by immunoprecipitation and analyzed by gel electrophoresis and fluorography. In vivo, myeloperoxidase was labeled initially as a 85-K glycosylated polypeptide (75 K after treatment with endo-beta-N-acetylglucosaminidase H). This polypeptide was soon processed to an 81-K intermediate and to smaller mature fragments of 60 K and 13 K within approximately 1 day. A minor portion of the precursor was converted to fragments of 40 K and 43 K. The pattern of labeled polypeptides of mature myeloperoxidase was similar to that of the enzyme purified from human leucocytes. The modifications of the polypeptide and of the oligosaccharide side chains in myeloperoxidase resembled those known to occur during the processing of lysosomal enzymes. In the absence or presence of dog pancreas membranes, myeloperoxidase was synthesized in vitro as a 76-K polypeptide or a 87-K glycosylated polypeptide, respectively. In HL-60 cells [32P]phosphate was incorporated into endo-beta-N-acetylglucosaminidase H-sensitive oligosaccharides. The presence of phosphorylated oligosaccharides was inferred from the fact that endocytosis of leucocyte myeloperoxidase in fibroblasts was sensitive to mannose 6-phosphate. It is suggested that myeloperoxidase is synthesized in the rough endoplasmic reticulum as a precursor of larger molecular mass and that the oligosaccharide side chains in the precursor are modified to contain mannose 6-phosphate residues which may be involved in the segregation and transport of the precursor.

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Selected References

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

  1. Andersen M. R., Atkin C. L., Eyre H. J. Intact form of myeloperoxidase from normal human neutrophils. Arch Biochem Biophys. 1982 Mar;214(1):273–283. doi: 10.1016/0003-9861(82)90031-5. [DOI] [PubMed] [Google Scholar]
  2. Collins S. J., Gallo R. C., Gallagher R. E. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature. 1977 Nov 24;270(5635):347–349. doi: 10.1038/270347a0. [DOI] [PubMed] [Google Scholar]
  3. Erickson A. H., Conner G. E., Blobel G. Biosynthesis of a lysosomal enzyme. Partial structure of two transient and functionally distinct NH2-terminal sequences in cathepsin D. J Biol Chem. 1981 Nov 10;256(21):11224–11231. [PubMed] [Google Scholar]
  4. Frisch A., Neufeld E. F. Limited proteolysis of the beta-hexosaminidase precursor in a cell-free system. J Biol Chem. 1981 Aug 10;256(15):8242–8246. [PubMed] [Google Scholar]
  5. GORHAM L. W., WAYMOUTH C. DIFFERENTIATION IN VITRO OF EMBRYONIC CARTILAGE AND BONE IN A CHEMICALLY-DEFINED MEDIUM. Proc Soc Exp Biol Med. 1965 May;119:287–290. doi: 10.3181/00379727-119-30160. [DOI] [PubMed] [Google Scholar]
  6. Gabel C. A., Goldberg D. E., Kornfeld S. 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):775–779. doi: 10.1073/pnas.80.3.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gieselmann V., Pohlmann R., Hasilik A., Von Figura K. Biosynthesis and transport of cathepsin D in cultured human fibroblasts. J Cell Biol. 1983 Jul;97(1):1–5. doi: 10.1083/jcb.97.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Hasilik A., Pohlmann R., von Figura K. Inhibition by cyanate of the processing of lysosomal enzymes. Biochem J. 1983 Mar 15;210(3):795–802. doi: 10.1042/bj2100795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hasilik A., Von Figura K. Oligosaccharides in lysosomal enzymes. Distribution of high-mannose and complex oligosaccharides in cathepsin D and beta-hexosaminidase. Eur J Biochem. 1981 Dec;121(1):125–129. doi: 10.1111/j.1432-1033.1981.tb06440.x. [DOI] [PubMed] [Google Scholar]
  11. Howard B. H., Adams S. L., Sobel M. E., Pastan I., de Crombrugghe B. Decreased levels of collagen mRNA in rous sarcoma virus-transformed chick embryo fibroblasts. J Biol Chem. 1978 Aug 25;253(16):5869–5874. [PubMed] [Google Scholar]
  12. Kimura S., Elce J. S., Jellinck P. H. Immunological relationship between peroxidases in eosinophils, uterus and other tissues of the rat. Biochem J. 1983 Jul 1;213(1):165–169. doi: 10.1042/bj2130165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Matheson N. R., Wong P. S., Travis J. Isolation and properties of human neutrophil myeloperoxidase. Biochemistry. 1981 Jan 20;20(2):325–330. doi: 10.1021/bi00505a015. [DOI] [PubMed] [Google Scholar]
  15. Olsen R. L., Little C. Purification and some properties of myeloperoxidase and eosinophil peroxidase from human blood. Biochem J. 1983 Mar 1;209(3):781–787. doi: 10.1042/bj2090781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Olsson I., Olofsson T. Induction of differentiation in a human promyelocytic leukemic cell line (HL-60). Production of granule proteins. Exp Cell Res. 1981 Jan;131(1):225–230. doi: 10.1016/0014-4827(81)90422-5. [DOI] [PubMed] [Google Scholar]
  17. Pember S. O., Shapira R., Kinkade J. M., Jr Multiple forms of myeloperoxidase from human neutrophilic granulocytes: evidence for differences in compartmentalization, enzymatic activity, and subunit structure. Arch Biochem Biophys. 1983 Mar;221(2):391–403. doi: 10.1016/0003-9861(83)90158-3. [DOI] [PubMed] [Google Scholar]
  18. Proia R. L., Neufeld E. F. Synthesis of beta-hexosaminidase in cell-free translation and in intact fibroblasts: an insoluble precursor alpha chain in a rare form of Tay-Sachs disease. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6360–6364. doi: 10.1073/pnas.79.20.6360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rosenfeld M. G., Kreibich G., Popov D., Kato K., Sabatini D. D. Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution. J Cell Biol. 1982 Apr;93(1):135–143. doi: 10.1083/jcb.93.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sly W. S., Fischer H. D. The phosphomannosyl recognition system for intracellular and intercellular transport of lysosomal enzymes. J Cell Biochem. 1982;18(1):67–85. doi: 10.1002/jcb.1982.240180107. [DOI] [PubMed] [Google Scholar]
  21. Ullrich K., Mersmann G., Weber E., Von Figura K. Evidence for lysosomal enzyme recognition by human fibroblasts via a phosphorylated carbohydrate moiety. Biochem J. 1978 Mar 15;170(3):643–650. doi: 10.1042/bj1700643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Warren G., Dobberstein B. Protein transfer across microsomal membranes reassembled from separated membrane components. Nature. 1978 Jun 15;273(5663):569–571. doi: 10.1038/273569a0. [DOI] [PubMed] [Google Scholar]
  23. Yamada M., Kurahashi K. Regulation of myeloperoxidase gene expression during differentiation of human myeloid leukemia HL-60 cells. J Biol Chem. 1984 Mar 10;259(5):3021–3025. [PubMed] [Google Scholar]
  24. Yamada M., Mori M., Sugimura T. Purification and characterization of small molecular weight myeloperoxidase from human promyelocytic leukemia HL-60 cells. Biochemistry. 1981 Feb 17;20(4):766–771. doi: 10.1021/bi00507a018. [DOI] [PubMed] [Google Scholar]
  25. von Figura K. Human alpha-N-acetylglucosaminidase. 1. Purification and properties. Eur J Biochem. 1977 Nov 1;80(2):523–533. [PubMed] [Google Scholar]

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