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. 1968 Mar 31;127(4):833–851. doi: 10.1084/jem.127.4.833

INTRAPHAGOCYTIC β-N-ACETYLGLUCOSAMINIDASE

PROPERTIES OF THE ENZYME AND ITS ACTIVITY ON GROUP A STREPTOCOCCAL CARBOHYDRATE IN COMPARISON WITH A SOIL BACILLUS ENZYME

Elia M Ayoub 1, Maclyn McCarty 1
PMCID: PMC2138474  PMID: 5642468

Abstract

The β-N-acetylglucosaminidases of rabbit and human polymorphonuclear leukocytes and of rabbit alveolar macrophages have been studied in comparison with the β-N-acetylglucosaminidase derived from a soil bacillus which had previously been shown to hydrolyze the group-specific polysaccharide of Group A streptococci. The phagocytic enzymes are lysosome associated and have an acid pH optimum. In contrast, the soil bacillus enzyme is an extracellular product, has a higher pH optimum, and is probaby of smaller molecular size. When tested on p-nitrophenyl-βN-acetylglucosaminide as substrate, the Km of the phagocytic enzymes is slightly higher than that of the soil bacillus. However, there were extreme differences in their effect on the Group A streptococcal polysaccharide. Thus, 5 x 106 units of the alveolar macrophage enzyme were required to hydrolyze the available N-acetylglucosamine of 1 mg of polysaccharide in 18 hr, while 100 units of the soil bacillus enzyme were sufficient to achieve this hydrolysis. In both cases, the serological reactivity of the polysaccharide is altered with loss of Group A specificity and acquisition of a new specificity characteristic of A-variant streptococci. Possible explanations for differences in the activity of the enzymes are considered, and the role of the phagocytic enzymes in intracellular degradation of Group A streptococci is discussed.

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Selected References

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  1. Ayoub E. M., Wannamaker L. W. The fate of group A streptococci following phagocytosis. In vitro phagocytic studies of isotope-labeled streptococci. J Immunol. 1967 Dec;99(6):1099–1105. [PubMed] [Google Scholar]
  2. BLEIWEIS A. S., KARAKAWA W. W., KRAUSE R. M. IMPROVED TECHNIQUE FOR THE PREPARATION OF STREPTOCOCCAL CELL WALLS. J Bacteriol. 1964 Oct;88:1198–1200. doi: 10.1128/jb.88.4.1198-1200.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COHN Z. A., HIRSCH J. G. The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. J Exp Med. 1960 Dec 1;112:983–1004. doi: 10.1084/jem.112.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. COHN Z. A., WIENER E. THE PARTICULATE HYDROLASES OF MACROPHAGES. I. COMPARATIVE ENZYMOLOGY, ISOLATION, AND PROPERTIES. J Exp Med. 1963 Dec 1;118:991–1008. doi: 10.1084/jem.118.6.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  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. FINDLAY J., LEVVY G. A. Purification of beta-N-acetylglucosaminidase from the pig epididymis. Biochem J. 1960 Oct;77:170–175. doi: 10.1042/bj0770170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HEYMANN H., MANNIELLO J. M., ZELEZNICK L. D., BARKULIS S. S. STRUCTURE OF STREPTOCOCCAL CELL WALLS. II. GROUP A BIOSE AND GROUP A TRIOSE FROM C-POLYSACCHARIDE. J Biol Chem. 1964 May;239:1656–1663. [PubMed] [Google Scholar]
  9. HUGHES R. C., JEANLOZ R. W. THE EXTRACELLULAR GLYCOSIDASES OF DIPLOCOCCUS PNEUMONIAE. II. PURIFICATION AND PROPERTIES OF A BETA-N-ACETYLGLUCOSAMINIDASE. ACTION ON A DERIVATIVE ON THE ALPHA-1-ACID GLYCOPROTEIN OF HUMAN PLASMA. Biochemistry. 1964 Oct;3:1543–1548. doi: 10.1021/bi00898a026. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. MCCARTY M. Variation in the group-specific carbohydrate of group A streptococci. II. Studies on the chemical basis for serological specificity of the carbohydrates. J Exp Med. 1956 Nov 1;104(5):629–643. doi: 10.1084/jem.104.5.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McCARTY M., LANCEFIELD R. C. Variation in the group-specific carbohydrate of group A streptococci. I. Immunochemical studies on the carbohydrates of variant strains. J Exp Med. 1955 Jul 1;102(1):11–28. doi: 10.1084/jem.102.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. PUGH D., LEABACK D. H., WALKER P. G. Studies on glucosaminidase; N-acetyl-beta-glucosaminidase in rat kidney. Biochem J. 1957 Mar;65(3):464–469. doi: 10.1042/bj0650464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. PUGH D., WALKER P. G. The localization of N-acetyl-beta-glucosaminidase in tissues. J Histochem Cytochem. 1961 May;9:242–250. doi: 10.1177/9.3.242. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. RONDLE C. J., MORGAN W. T. The determination of glucosamine and galactosamine. Biochem J. 1955 Dec;61(4):586–589. doi: 10.1042/bj0610586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SALTON M. R. J., HORNE R. W. Studies of the bacterial cell wall. II. Methods of preparation and some properties of cell walls. Biochim Biophys Acta. 1951 Jul;7(2):177–197. doi: 10.1016/0006-3002(51)90017-0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Schwab J. H., Ohanian S. H. Degradation of streptococcal cell wall antigens in vivo. J Bacteriol. 1967 Nov;94(5):1346–1352. doi: 10.1128/jb.94.5.1346-1352.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Swift H. F., Wilson A. T., Lancefield R. C. TYPING GROUP A HEMOLYTIC STREPTOCOCCI BY M PRECIPITIN REACTIONS IN CAPILLARY PIPETTES. J Exp Med. 1943 Aug 1;78(2):127–133. doi: 10.1084/jem.78.2.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. WATKINS W. M. Enzymes of Trichomonas foetus; the action of cell-free extracts on blood-group substances and low-molecular-weight glycosides. Biochem J. 1959 Feb;71(2):261–274. doi: 10.1042/bj0710261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. WOOLLEN J. W., HEYWORTH R., WALKER P. G. Studies on glucosaminidase. 3. Testicular N-acetyl-beta-glucosaminidase and N-acetyl-beta-galactosaminidase. Biochem J. 1961 Jan;78:111–116. doi: 10.1042/bj0780111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. WOOLLEN J. W., WALKER P. G., HEYWORTH R. Studies on glucosaminidase. 6. N-Acetyl-beta-glucosaminidase and N-acetyl-beta-galactosaminidase activities of a variety of enzyme preparations. Biochem J. 1961 May;79:294–298. doi: 10.1042/bj0790294. [DOI] [PMC free article] [PubMed] [Google Scholar]

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