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. 1973 Jun 1;137(6):1459–1471. doi: 10.1084/jem.137.6.1459

IDENTIFICATION OF A MAJOR BASIC PROTEIN IN GUINEA PIG EOSINOPHIL GRANULES

Gerald J Gleich 1, David A Loegering 1, Jorge E Maldonado 1
PMCID: PMC2139342  PMID: 4350835

Abstract

Elucidation of the functions of the eosinophil might be accomplished by analysis of the granule constituents. We have purified eosinophils (93% or greater) from the peritoneal cavity of the guinea pig and have investigated a variety of methods to disrupt cells and liberate intact granules. Lysis in 0.34 M sucrose gave the best yield of granules and these had the characteristic morphology of eosinophil granules when examined by electron microscopy. Granules were solubilized by a variety of treatments and the solutions analyzed by polyacrylamide electrophoresis at pH 3 in 6 M urea. Comparison of the electrophoretic patterns of solubilized eosinophil and neutrophil granules revealed a difference: a major portion (53±3%; x ±1 SE) of the protein from the eosinophil granule migrated as a single component. This major band protein has a molecular weight between 6,000 and 12,000 daltons and a pI of 10 or greater. Analysis of eosinophil granule constituents on Sephadex G-50 revealed two main peaks; peak 1 possessed peroxidase activity and peak 2 contained the major band protein. These studies indicate that eosinophil granules contain a cationic protein of low molecular weight which lacks peroxidase activity and which accounts for greater than 50% of granule protein.

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Selected References

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  1. ARCHER G. T., HIRSCH J. G. ISOLATION OF GRANULES FROM EOSINOPHIL LEUCOCYTES AND STUDY OF THEIR ENZYME CONTENT. J Exp Med. 1963 Aug 1;118:277–286. doi: 10.1084/jem.118.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Cotran R. S., Litt M. The entry of granule-associated peroxidase into the phagocytic vacuoles of eosinophils. J Exp Med. 1969 Jun 1;129(6):1291–1306. doi: 10.1084/jem.129.6.1291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
  5. Desser R. K., Himmelhoch S. R., Evans W. H., Januska M., Mage M., Shelton E. Guinea pig heterophil and eosinophil peroxidase. Arch Biochem Biophys. 1972 Feb;148(2):452–465. doi: 10.1016/0003-9861(72)90164-6. [DOI] [PubMed] [Google Scholar]
  6. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  7. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. LITT M. Studies in experimental eosinophilia. I. Repeated quantitation of peritoneal eosinophilia in guinea pigs by a method of peritoneal lavage. Blood. 1960 Sep;16:1318–1329. [PubMed] [Google Scholar]
  10. PEASE D. C. An electron microscopic study of red bone marrow. Blood. 1956 Jun;11(6):501–526. [PubMed] [Google Scholar]
  11. Panyim S., Chalkley R. High resolution acrylamide gel electrophoresis of histones. Arch Biochem Biophys. 1969 Mar;130(1):337–346. doi: 10.1016/0003-9861(69)90042-3. [DOI] [PubMed] [Google Scholar]
  12. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  13. Ranadive N. S., Cochrane C. G. Isolation and characterization of permeability factors from rabbit neutrophils. J Exp Med. 1968 Oct 1;128(4):605–622. doi: 10.1084/jem.128.4.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Seegers W., Janoff A. Mediators of inflammation in leukocyte lysosomes. VI. Partial purification and characterization of a mast cell-rupturing component. J Exp Med. 1966 Nov 1;124(5):833–849. doi: 10.1084/jem.124.5.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. VERCAUTEREN R., PEETERS G. On the presence of an antihistaminicum in isolated eosinophilic granulocytes. Arch Int Pharmacodyn Ther. 1952 Feb;89(1):10–14. [PubMed] [Google Scholar]
  16. VERCAUTEREN R. The properties of the isolated granules from blood eosinophiles. Enzymologia. 1953 Apr 15;16(1):1–13. [PubMed] [Google Scholar]
  17. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  18. 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]
  19. 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]

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