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. 1966 Feb;91(2):755–762. doi: 10.1128/jb.91.2.755-762.1966

Cationic Proteins of Polymorphonuclear Leukocyte Lysosomes II. Composition, Properties, and Mechanism of Antibacterial Action

H I Zeya a, J K Spitznagel a,1
PMCID: PMC314925  PMID: 5934974

Abstract

Zeya, H. I. (University of North Carolina, Chapel Hill), and J. K. Spitznagel. Cationic proteins of polymorphonuclear leukocyte lysosomes. II. Composition, properties, and mechanism of antibacterial action. J. Bacteriol. 91:755–762. 1966.—A basic proteins fraction from guinea pig polymorphonuclear (PMN) granules was obtained by acid extraction and precipitation with 20% (v/v) ethyl alcohol. The fraction accounted for most of the antibacterial activity of the PMN granules and corresponded to the antibacterial cationic components of intact granules (bands I, II, and III) resolved by zone electrophoresis. Absence from the fraction of components identical to the enzymatic components of intact lysosomes showed that the fraction was essentially free from lysosomal enzymes. The amino acid analysis of proteins in the fraction gave a preponderance of basic amino acids (25%), especially of arginine (16%). The comparative amino acid analysis showed that the lysosomal cationic proteins (LCP) fraction was markedly different from nuclear histones. The LCP fraction manifested antibacterial activity against certain gram-positive and gram-positive microorganisms, including Candida albicans, and exhibited stoichiometric relationship in its activity. Microorganisms treated with LCP fraction were agglutinated. Anionic substances such as nucleic acids, heparin, and endotoxin effectively blocked the antibacterial activity of the fraction. The LCP fraction caused suppression of oxygen uptake by bacterial cells and damaged the permeability barriers of cells as manifested by rapid release of P32 as well as ultraviolet-absorbing material at 260 mμ, in the supernatant fluid.

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Selected References

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  1. BECKER F. F., GREEN H. The effects of protamines and histones on the nucleic acids of ascites tumor cells. Exp Cell Res. 1960 Mar;19:361–375. doi: 10.1016/0014-4827(60)90014-8. [DOI] [PubMed] [Google Scholar]
  2. BLOOM W. L., WINTERS M. G., WATSON D. W. The inhibition of two antibacterial basic proteins by nucleic acids. J Bacteriol. 1951 Jul;62(1):7–13. doi: 10.1128/jb.62.1.7-13.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BURGER W. C., STAHMANN M. A. The agglutination and growth inhibition of bacteria by lysine polypeptides. Arch Biochem Biophys. 1952 Jul;39(1):27–36. doi: 10.1016/0003-9861(52)90257-9. [DOI] [PubMed] [Google Scholar]
  4. BUTLER J. A., COHN P., SIMSON P. The presence of basic proteins in microsomes. Biochim Biophys Acta. 1960 Feb 26;38:386–388. doi: 10.1016/0006-3002(60)91272-5. [DOI] [PubMed] [Google Scholar]
  5. BUTLER J. A., DAVISON P. F., JAMES D. W. Isolation and properties of histones from thymus nucleoproteins. Biochem J. 1953 Jun;54(3):xxi–xxi. [PubMed] [Google Scholar]
  6. BUTLER J. A., DAVISON P. F., JAMES D. W., SHOOTER K. V. The histones of calf thymus deoxyribonucleoprotein. I. Preparation and homogeneity. Biochim Biophys Acta. 1954 Feb;13(2):224–232. doi: 10.1016/0006-3002(54)90307-8. [DOI] [PubMed] [Google Scholar]
  7. CIECIURA S. J., MARCUS P. I., PUCK T. T. Clonal growth in vitro of epithelial cells from normal human tissues. J Exp Med. 1956 Oct 1;104(4):615–628. doi: 10.1084/jem.104.4.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. CRAMPTON C. F., MOORE S., STEIN W. H. Chromatographic fractionation of calf thymus histone. J Biol Chem. 1955 Aug;215(2):787–801. [PubMed] [Google Scholar]
  10. DALY M. M., MIRSKY A. E. Histones with high lysine content. J Gen Physiol. 1955 Jan 20;38(3):405–413. doi: 10.1085/jgp.38.3.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. DAVISON P. F., BUTLER J. A. The fractionation and composition of histones from thymus nucleoprotein. Biochim Biophys Acta. 1954 Nov;15(3):439–440. doi: 10.1016/0006-3002(54)90051-7. [DOI] [PubMed] [Google Scholar]
  12. HIRSCH J. G. Bactericidal action of histone. J Exp Med. 1958 Dec 1;108(6):925–944. doi: 10.1084/jem.108.6.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. HIRSCH J. G. Cinemicrophotographic observations on granule lysis in polymorphonuclear leucocytes during phagocytosis. J Exp Med. 1962 Dec 1;116:827–834. doi: 10.1084/jem.116.6.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. HORN R. G., SPICER S. S., WETZEL B. K. PHAGOCYTOSIS OF BACTERIA BY HETEROPHIL LEUKOCYTES: ACID AND ALKALINE PHOSPHATASE CYTOCHEMISTRY. Am J Pathol. 1964 Aug;45:327–335. [PMC free article] [PubMed] [Google Scholar]
  15. JANOFF A., ZWEIFACH B. W. PRODUCTION OF INFLAMMATORY CHANGES IN THE MICROCIRCULATION BY CATIONIC PROTEINS EXTRACTED FROM LYSOSOMES. J Exp Med. 1964 Nov 1;120:747–764. doi: 10.1084/jem.120.5.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. KATCHALSKI E., BICHOWSKI-SLOMNITZKI L., VOLCANI B. E. The action of some water-soluble poly-alpha-amino acids on bacteria. Biochem J. 1953 Nov;55(4):671–680. doi: 10.1042/bj0550671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LESLIE I. Biochemistry of heredity: a general hypothesis. Nature. 1961 Jan 28;189:260–268. doi: 10.1038/189260a0. [DOI] [PubMed] [Google Scholar]
  18. Miller B. F., Abrams R., Dorfman A., Klein M. ANTIBACTERIAL PROPERTIES OF PROTAMINE AND HISTONE. Science. 1942 Nov 6;96(2497):428–430. doi: 10.1126/science.96.2497.428. [DOI] [PubMed] [Google Scholar]
  19. NEWTON B. A. The properties and mode of action of the polymyxins. Bacteriol Rev. 1956 Mar;20(1):14–27. doi: 10.1128/br.20.1.14-27.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. SALTON M. R. J. The adsorption of cetyltrimethylammonium bromide by bacteria, its action in releasing cellular constituents and its bactericidal effects. J Gen Microbiol. 1951 May;5(2):391–404. doi: 10.1099/00221287-5-2-391. [DOI] [PubMed] [Google Scholar]
  21. SKARNES R. C., WATSON D. W. Characterization of leukin: an antibacterial factor from leucocytes active against gram-positive pathogens. J Exp Med. 1956 Dec 1;104(6):829–845. doi: 10.1084/jem.104.6.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SPECTOR W. G., WILLOUGHBY D. A. The inflammatory response. Bacteriol Rev. 1963 Jun;27:117–154. doi: 10.1128/br.27.2.117-154.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. SPITZNAGEL J. K., CHI H. Y. CATIONIC PROTEINS AND ANTIBACTERIAL PROPERTIES OF INFECTED TISSUES AND LEUKOCYTES. Am J Pathol. 1963 Oct;43:697–711. [PMC free article] [PubMed] [Google Scholar]
  24. SPITZNAGEL J. K. The effects of mammalian and other cationic polypeptides on the cytochemical character of bacterial cells. J Exp Med. 1961 Dec 1;114:1063–1078. doi: 10.1084/jem.114.6.1063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. UI N. Preparation, fractionation and properties of calf thymus histone. Biochim Biophys Acta. 1957 Sep;25(3):493–502. doi: 10.1016/0006-3002(57)90519-x. [DOI] [PubMed] [Google Scholar]
  26. WEISSMANN G., BECHER B., WIEDERMANN G., BERNHEIMER A. W. STUDIES ON LYSOSOMES. VII. ACUTE AND CHRONIC ARTHRITIS PRODUCED BY INTRA-ARTICULAR INJECTIONS OF STREPTOLYSIN S IN RABBITS. Am J Pathol. 1965 Jan;46:129–147. [PMC free article] [PubMed] [Google Scholar]
  27. ZEYA H. I., SPITZNAGEL J. K. ANTIBACTERIAL AND ENZYMIC BASIC PROTEINS FROM LEUKOCYTE LYSOSOMES: SEPARATION AND IDENTIFICATION. Science. 1963 Nov 22;142(3595):1085–1087. doi: 10.1126/science.142.3595.1085. [DOI] [PubMed] [Google Scholar]
  28. Zeya H. I., Spitznagel J. K. Cationic proteins of polymorphonuclear leukocyte lysosomes. I. Resolution of antibacterial and enzymatic activities. J Bacteriol. 1966 Feb;91(2):750–754. doi: 10.1128/jb.91.2.750-754.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]

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