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. 1965 Nov;97(2):380–388. doi: 10.1042/bj0970380

Studies on bone enzymes. The assay of acid hydrolases and other enzymes in bone tissue

G Vaes 1, P Jacques 1
PMCID: PMC1264652  PMID: 16749142

Abstract

1. Nine acid hydrolases, cytochrome oxidase, alkaline phenylphosphatase and catalase were demonstrated in 0·25m-sucrose homogenates of newborn-rat calvaria. The acid hydrolases were: acid phenylphosphatase, acid β-glycerophosphatase, β-glucuronidase, β-N-acetylglucosaminidase (β-N-acetylaminodeoxyglucosidase), acid ribonuclease and acid deoxyribonuclease, showing optimum activity at about pH5; cathepsin, β-galactosidase and hyaluronidase, with optimum activity at about pH3·6. 2. The main kinetic characters of these enzymes have been studied and methods for their quantitative assay have been worked out. The activities present in bone are given and compared with those found in liver. 3. Acid-phosphatase activity was assayed with phenyl phosphate and β-glycerophosphate as substrates: activities with these two substrates appeared to be due to two different enzymes. Acid phenylphosphatase is particularly labile and is readily inactivated by various physical or chemical agents.

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Selected References

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

  1. APPELMANS F., DE DUVE C. Tissue fractionation studies. 3. Further observations on the binding of acid phosphatase by rat-liver particles. Biochem J. 1955 Mar;59(3):426–433. doi: 10.1042/bj0590426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. APPELMANS F., WATTIAUX R., DE DUVE C. Tissue fractionation studies. 5. The association of acid phosphatase with a special class of cytoplasmic granules in rat liver. Biochem J. 1955 Mar;59(3):438–445. doi: 10.1042/bj0590438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BORLE A. B., NICHOLS N., NICHOLS G., Jr Metabolic studies of bone in vitro. II. The metabolic patterns of accretion and resorption. J Biol Chem. 1960 Apr;235:1211–1214. [PubMed] [Google Scholar]
  4. Baudhuin P., Beaufay H., Rahman-Li Y., Sellinger O. Z., Wattiaux R., Jacques P., De Duve C. Tissue fractionation studies. 17. Intracellular distribution of monoamine oxidase, aspartate aminotransferase, alanine aminotransferase, D-amino acid oxidase and catalase in rat-liver tissue. Biochem J. 1964 Jul;92(1):179–184. doi: 10.1042/bj0920179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CAMERON D. A. The fine structure of bone and calcified cartilage. A critical review of the contribution of electron microscopy to the understading of osteogenesis. Clin Orthop Relat Res. 1963;26:199–228. [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. 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]
  9. HOLT S. J., HICKS R. M. Studies on formalin fixation for electron microscopy and cytochemical staining purposes. J Biophys Biochem Cytol. 1961 Oct;11:31–45. doi: 10.1083/jcb.11.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. KIND P. R., KING E. J. Estimation of plasma phosphatase by determination of hydrolysed phenol with amino-antipyrine. J Clin Pathol. 1954 Nov;7(4):322–326. doi: 10.1136/jcp.7.4.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. NORDIN B. E. [The solubility of powdered bone]. J Biol Chem. 1957 Aug;227(2):551–564. [PubMed] [Google Scholar]
  13. PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
  14. REISSIG J. L., STORMINGER J. L., LELOIR L. F. A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem. 1955 Dec;217(2):959–966. [PubMed] [Google Scholar]
  15. SCHARTUM S., NICHOLS G., Jr Concerning pH gradients between the extracellular compartment and fluids bathing the bone mineral surface and their relation to calcium ion distribution. J Clin Invest. 1962 May;41:1163–1168. doi: 10.1172/JCI104569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. SCHLAGER F. [Occurrence and localization of beta-D-galactosidase in bone, cartilage and adjacent tissue of white mice]. Acta Histochem. 1959 Oct 13;8:176–184. [PubMed] [Google Scholar]
  17. 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]
  18. TOLKSDORF S. The in vitro determination of hyaluronidase. Methods Biochem Anal. 1954;1:425–457. doi: 10.1002/9780470110171.ch15. [DOI] [PubMed] [Google Scholar]
  19. TONNA E. A. Histologic and histochemical studies on the periosteum of male and female rats at different ages. J Gerontol. 1958 Jan;13(1):14–19. doi: 10.1093/geronj/13.1.14. [DOI] [PubMed] [Google Scholar]
  20. VAES G. M., NICHOLS G., Jr Effects of a dose of parathyroid extract on bone metabolic pathways. Endocrinology. 1962 Apr;70:546–555. doi: 10.1210/endo-70-4-546. [DOI] [PubMed] [Google Scholar]
  21. Vaes G., Jacques P. Studies on bone enzymes. Distribution of acid hydrolases, alkaline phenylphosphatase, cytochrome oxidase and catalase in subcellular fraction of bone tissue homogenates. Biochem J. 1965 Nov;97(2):389–392. doi: 10.1042/bj0970389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vaes G. Studies on bone enzymes. The activation and release of latent acid hydrolases and catalase in bone-tissue homogenates. Biochem J. 1965 Nov;97(2):393–402. doi: 10.1042/bj0970393. [DOI] [PMC free article] [PubMed] [Google Scholar]

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