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. 1967 Feb 1;32(2):325–337. doi: 10.1083/jcb.32.2.325

LYSOSOMES IN LYMPHOID TISSUE

I. The Measurement of Hydrolytic Activities in Whole Homogenates

William E Bowers 1, John T Finkenstaedt 1, Christian de Duve 1
PMCID: PMC2107247  PMID: 10976225

Abstract

Methods have been developed for the quantitative assay of cytochrome oxidase, esterase, and 11 acid hydrolases in rat-spleen homogenates. These methods seem to be applicable also to other lymphoid tissues. Preliminary studies, extended to nine of the acid hydrolases, indicate that these enzymes occur in partly latent and sedimentable form and that they can be unmasked and rendered soluble by some of the treatments that liberate the enzymes from rat-liver lysosomes. The spleen particles appear to be very sensitive to mechanical injury, a property which necessitates special precautions in homogenizing the tissue. Agglutination of spleen particles takes place to a larger extent in 0.25 M sucrose than in 0.15 M KCl.

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Selected References

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

  1. BEAUFAY H., DE DUVE C., HOLT S. J., UNDERHAY E. Intracellular localization of esterase in rat liver. J Biophys Biochem Cytol. 1956 Sep 25;2(5):635–637. doi: 10.1083/jcb.2.5.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BOUMA J. M., GRUBER M. THE DISTRIBUTION OF CATHEPSINS B AND C IN RAT TISSUES. Biochim Biophys Acta. 1964 Sep 18;89:545–547. doi: 10.1016/0926-6569(64)90082-3. [DOI] [PubMed] [Google Scholar]
  3. Bouma J. M., Gruber M. Intracellular distribution of cathepsin B and cathepsin C in rat liver. Biochim Biophys Acta. 1966 Feb 14;113(2):350–358. doi: 10.1016/s0926-6593(66)80074-7. [DOI] [PubMed] [Google Scholar]
  4. CONCHIE J., FINDLAY J., LEVVY G. A. Mammalian glycosidases; distribution in the body. Biochem J. 1959 Feb;71(2):318–325. doi: 10.1042/bj0710318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CONCHIE J., HAY A. J., LEVVY G. A. Mammalian glycosidases. 3. The intracellular localization of beta-glucuronidase in different mammalian tissues. Biochem J. 1961 May;79:324–330. doi: 10.1042/bj0790324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CONCHIE J., HAY A. J. Mammalian glycosidases. 4. The intracellular localization of beta-galactosidase, alpha-mannosidase, beta-N-acetylglucosaminidase and alpha-L-fucosidase in mammalian tissues. Biochem J. 1963 May;87:354–361. doi: 10.1042/bj0870354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. COOPERSTEIN S. J., LAZAROW A. A microspectrophotometric method for the determination of cytochrome oxidase. J Biol Chem. 1951 Apr;189(2):665–670. [PubMed] [Google Scholar]
  8. DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DOUNCE A. L., WITTER R. F., MONTY K. J., PATE S., COTTONE M. A. A method for isolating intact mitochondria and nuclei from the same homogenate, and the influence of mitochondrial destruction on the properties of cell nuclei. J Biophys Biochem Cytol. 1955 Mar;1(2):139–153. doi: 10.1083/jcb.1.2.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. EICHEL H. J., ROTH J. S. Intracellular localization of enzymes in spleen. II. Some properties and the distribution of ribonuclease in rat spleen. J Cell Biol. 1962 Feb;12:263–276. doi: 10.1083/jcb.12.2.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. FINDLAY J., LEVVY G. A., MARSH C. A. Inhibition of glycosidases by aldonolactones of corresponding configuration. 2. Inhibitors of beta-N-acetylglucosaminidase. Biochem J. 1958 Jul;69(3):467–476. doi: 10.1042/bj0690467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. GIANETTO R., DE DUVE C. Tissue fractionation studies. 4. Comparative study of the binding of acid phosphatase, beta-glucuronidase and cathepsin by rat-liver particles. Biochem J. 1955 Mar;59(3):433–438. doi: 10.1042/bj0590433. [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. RAHMAN Y. E. A NOTE ON ACID PHOSPHATASE RELEASE FROM SPLEEN, LIVER AND THYMUS OF RATS. Biochim Biophys Acta. 1964 Aug 19;90:440–442. doi: 10.1016/0304-4165(64)90221-1. [DOI] [PubMed] [Google Scholar]
  15. RAHMAN Y. E. Electron microscopy of lysosome-rich fractions from rat thymus isolated by density-gradient centrifugation before and after whole-body x-irradiation. J Cell Biol. 1962 May;13:253–260. doi: 10.1083/jcb.13.2.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. ROTH J. S., EICHEL H. J. The effect of total-body x-irradiation on the distribution of ribonuclease activity in subcellular fractions of rat spleen. Radiat Res. 1959 Oct;11:572–581. [PubMed] [Google Scholar]
  17. ROY A. B. Comparative studies on the liver sulphatases. Biochem J. 1958 Mar;68(3):519–528. doi: 10.1042/bj0680519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SACHS G., DEDUVE C., DVORKIN B. S., WHITE A. Effect of adrenal cortical steroid injection on lysosmal enzymic activities of rat thymus. Exp Cell Res. 1962 Dec;28:597–600. doi: 10.1016/0014-4827(62)90267-7. [DOI] [PubMed] [Google Scholar]
  19. SELIGSON D., HIRAHARA K. The measurement of ammonia in whole blood, erythrocytes, and plasma. J Lab Clin Med. 1957 Jun;49(6):962–974. [PubMed] [Google Scholar]
  20. SELLINGER O. Z., BEAUFAY H., JACQUES P., DOYEN A., DE DUVE C. Tissue fractionation studies. 15. Intracellular distribution and properties of beta-N-acetylglucosaminidase and beta-galactosidase in rat liver. Biochem J. 1960 Mar;74:450–456. doi: 10.1042/bj0740450. [DOI] [PMC free article] [PubMed] [Google Scholar]

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