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. 1978 Jul 15;174(1):53–59. doi: 10.1042/bj1740053

Subcellular distribution of glycosidases in human polymorphonuclear leucocytes.

R F Rest, M H Cooney, J K Spitznagel
PMCID: PMC1185884  PMID: 29608

Abstract

The subcellular distribution of nine glycosidases were studied in fractions of homogenized human polymorphonuclear leucocytes (neutrophils) obtained by isopycnic centrifugation through linear sucrose density gradients. The substrates were 4-methylumbelliferyl glycosides. All nine glycosides were hydrolysed by enzymes in neutrophil cytosol fractions, and by enzymes in at least one granule population. alpha-Glucosidase activity sedimented in sucrose density gradients to a point (p = 1.180 g/ml) just above the specific granules, possibly the 'tertiary' granule population. The peak corresponding to alpha-glucosidase did not co-sediment with, but considerably overlapped, the peak corresponding to lactoferrin, a marker for specific granules (p = 1.187 g/ml). alpha-Galactosidase activity was found primarily in heavy azurophil granules (p = 1.222 g/ml). alpha-Mannosidase activity was found primarily in light azurophil granules (p = 1.206 g/ml), following the distribution of myeloperoxidase, the commonly used azurophil granule marker. beta-Glucosidase activity was concentrated in mitochondrial fractions (p = 1.160 g/ml). All other glycosidases presented complex distributions, with activities not restricted to one granule class. Granule-associated glycosidase activities were increased 2--38 times when measured in the presence of 0.05% Triton X-100, indicating latency of the enzymes within granules.

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Selected References

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

  1. Avila J. L., Convit J. Studies on human polymorphonuclear leukocyte enzymes. I. Assay of acid hydrolases and other enzymes. Biochim Biophys Acta. 1973 Feb 15;293(2):397–408. doi: 10.1016/0005-2744(73)90347-1. [DOI] [PubMed] [Google Scholar]
  2. Avila J. L., Convit J. Studies on human polymorphonuclear leukocyte enzymes. II. Comparative study of the physical properties of primary and specific granules. Biochim Biophys Acta. 1973 Feb 15;293(2):409–423. doi: 10.1016/0005-2744(73)90348-3. [DOI] [PubMed] [Google Scholar]
  3. BRIDGES R. A., BERENDES H., GOOD R. A. A fatal granulomatous disease of childhood; the clinical, pathological, and laboratory features of a new syndrome. AMA J Dis Child. 1959 Apr;97(4):387–408. [PubMed] [Google Scholar]
  4. Bainton D. F., Farquhar M. G. Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. II. Cytochemistry and electron microscopy of bone marrow cells. J Cell Biol. 1968 Nov;39(2):299–317. doi: 10.1083/jcb.39.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bretz U., Baggiolini M. Biochemical and morphological characterization of azurophil and specific granules of human neutrophilic polymorphonuclear leukocytes. J Cell Biol. 1974 Oct;63(1):251–269. doi: 10.1083/jcb.63.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HERSH E. M., BODEY G. P., NIES B. A., FREIREICH E. J. CAUSES OF DEATH IN ACUTE LEUKEMIA: A TEN-YEAR STUDY OF 414 PATIENTS FROM 1954-1963. JAMA. 1965 Jul 12;193:105–109. doi: 10.1001/jama.1965.03090020019005. [DOI] [PubMed] [Google Scholar]
  7. Kane S. P., Peters T. J. Analytical subcellular fractionation of human granulocytes with reference to the localization of vitamin B12-binding proteins. Clin Sci Mol Med. 1975 Aug;49(2):171–182. doi: 10.1042/cs0490171. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Leffell M. S., Spitznagel J. K. Association of lactoferrin with lysozyme in granules of human polymorphonuclear leukocytes. Infect Immun. 1972 Nov;6(5):761–765. doi: 10.1128/iai.6.5.761-765.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Leffell M. S., Spitznagel J. K. Fate of human lactoferrin and myeloperoxidase in phagocytizing human neutrophils: effects of immunoglobulin G subclasses and immune complexes coated on latex beads. Infect Immun. 1975 Oct;12(4):813–820. doi: 10.1128/iai.12.4.813-820.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  12. Querinjean P., Masson P. L., Heremans J. F. Molecular weight, single-chain structure and amino acid composition of human lactoferrin. Eur J Biochem. 1971 Jun 11;20(3):420–425. doi: 10.1111/j.1432-1033.1971.tb01408.x. [DOI] [PubMed] [Google Scholar]
  13. Rest R. F., Spitznagel J. K. Subcellular distribution of superoxide dismutases in human neutrophils. Influence of myeloperoxidase on the measurement of superoxide dismutase activity. Biochem J. 1977 Aug 15;166(2):145–153. doi: 10.1042/bj1660145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Spitznagel J. K., Dalldorf F. G., Leffell M. S., Folds J. D., Welsh I. R., Cooney M. H., Martin L. E. Character of azurophil and specific granules purified from human polymorphonuclear leukocytes. Lab Invest. 1974 Jun;30(6):774–785. [PubMed] [Google Scholar]
  15. West B. C., Rosenthal A. S., Gelb N. A., Kimball H. R. Separation and characterization of human neutrophil granules. Am J Pathol. 1974 Oct;77(1):41–66. [PMC free article] [PubMed] [Google Scholar]
  16. White J. G., Estensen R. D. Selective labilization of specific granules in polymorphonuclear leukocytes by phorbol myristate acetate. Am J Pathol. 1974 Apr;75(1):45–60. [PMC free article] [PubMed] [Google Scholar]
  17. Wright D. G., Bralove D. A., Gallin J. I. The differential mobilization of human neutrophil granules. Effects of phorbol myristate acetate and ionophore A23187. Am J Pathol. 1977 May;87(2):273–284. [PMC free article] [PubMed] [Google Scholar]

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