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. 1984 Jul;45(1):150–154. doi: 10.1128/iai.45.1.150-154.1984

Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes.

M E Selsted, D Szklarek, R I Lehrer
PMCID: PMC263292  PMID: 6735465

Abstract

Six antimicrobial peptides, corresponding to the family of "lysosomal cationic proteins" described previously by Zeya and Spitznagel (H. I. Zeya and J. K. Spitznagel, J. Bacteriol. 91:750-754, 1966; H. I. Zeya and J. K. Spitznagel, J. Bacteriol. 91:755-762, 1966), were purified from rabbit peritoneal granulocytes by preparative acrylamide gel electrophoresis and reversed-phase high-pressure liquid chromatography. Each of the peptides was of low molecular weight (ca. 4,000) as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The two most cationic peptides, NP-1 and NP-2, were active against a broad spectrum of gram-positive and gram-negative bacteria. The remaining four peptides, NP-3A, NP-3B, NP-4, and NP-5, had more selective antibacterial activity. None of the peptides was active against Bordetella bronchiseptica, a common pathogen of domestic rabbits. Antibacterial activity was best expressed at near neutral pH under conditions of low ionic strength.

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Selected References

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

  1. 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]
  2. Cech P., Lehrer R. I. Phagolysosomal pH of human neutrophils. Blood. 1984 Jan;63(1):88–95. [PubMed] [Google Scholar]
  3. HIRSCH J. G. Antimicrobial factors in tissues and phagocytic cells. Bacteriol Rev. 1960 Mar;24(1):133–140. doi: 10.1128/br.24.1.133-140.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. HIRSCH J. G. Phagocytin: a bactericidal substance from polymorphonuclear leucocytes. J Exp Med. 1956 May 1;103(5):589–611. doi: 10.1084/jem.103.5.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Lehrer R. I., Selsted M. E., Szklarek D., Fleischmann J. Antibacterial activity of microbicidal cationic proteins 1 and 2, natural peptide antibiotics of rabbit lung macrophages. Infect Immun. 1983 Oct;42(1):10–14. doi: 10.1128/iai.42.1.10-14.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Macrae E. K., Spitznagel J. K. Ultrastructural localization of cationic proteins in cytoplasmic granules of chicken and rabbit polymorphonuclear leukocytes. J Cell Sci. 1975 Jan;17(1):79–94. doi: 10.1242/jcs.17.1.79. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. 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]
  10. Segal A. W., Geisow M., Garcia R., Harper A., Miller R. The respiratory burst of phagocytic cells is associated with a rise in vacuolar pH. Nature. 1981 Apr 2;290(5805):406–409. doi: 10.1038/290406a0. [DOI] [PubMed] [Google Scholar]
  11. Selsted M. E., Brown D. M., DeLange R. J., Lehrer R. I. Primary structures of MCP-1 and MCP-2, natural peptide antibiotics of rabbit lung macrophages. J Biol Chem. 1983 Dec 10;258(23):14485–14489. [PubMed] [Google Scholar]
  12. 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]
  13. Zeya H. I., Spitznagel J. K. Arginine-rich proteins of polymorphonuclear leukocyte lysosomes. Antimicrobial specificity and biochemical heterogeneity. J Exp Med. 1968 May 1;127(5):927–941. doi: 10.1084/jem.127.5.927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Zeya H. I., Spitznagel J. K. Cationic protein-bearing granules of polymorphonuclear leukocytes: separation from enzyme-rich granules. Science. 1969 Mar 7;163(3871):1069–1071. doi: 10.1126/science.163.3871.1069. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Zeya H. I., Spitznagel J. K. Cationic proteins of polymorphonuclear leukocyte lysosomes. II. Composition, properties, and mechanism of antibacterial action. J Bacteriol. 1966 Feb;91(2):755–762. doi: 10.1128/jb.91.2.755-762.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Zeya H. I., Spitznagel J. K. Characterization of cationic protein-bearing granules of polymorphonuclear leukocytes. Lab Invest. 1971 Mar;24(3):229–236. [PubMed] [Google Scholar]

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