Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1987 Jun 15;244(3):675–682. doi: 10.1042/bj2440675

Identification and isolation of soluble histones from bovine milk and serum.

S Waga 1, E M Tan 1, R L Rubin 1
PMCID: PMC1148049  PMID: 3446184

Abstract

An immunoassay for soluble histones as trace components of biological fluids was developed on the basis of the dual capacity of histones to bind solid-phase DNA and monoclonal anti-histone antibody. Application of this histone-capture assay to bovine milk resulted in a positive signal, and DNA-cellulose chromatography was used to isolate histone-like material in microgram quantities. Western-blot analysis using a panel of anti-histone antibodies demonstrated the presence of histones H2A, H2B and H4 in apparently intact form. DNAase digestion experiments indicated that at least a portion of milk histone was complexed to DNA. Bovine serum was analysed in the same manner on serial DNA-cellulose columns, and H4 and partially degraded H2A were detected by Western-blot analysis. The finding of soluble histones in bovine milk and serum may account for unexpected results when these biological fluids are used as blocking reagents in Western blots and other immunoassays and may have ramifications in the origin and significance of anti-histone antibodies in human disease.

Full text

PDF
675

Images in this article

677Fig. 1.
on p.677
678Fig. 5.
on p.678
679Fig. 6.
on p.679
680Fig. 8.
on p.680
680Fig. 9.
on p.680

Selected References

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

  1. Anderson N. G., Powers M. T., Tollaksen S. L. Proteins of human milk. I. Identification of major components. Clin Chem. 1982 Apr;28(4 Pt 2):1045–1055. [PubMed] [Google Scholar]
  2. Atkinson M. J., Bell D. A., Singhal S. K. A naturally occurring polyclonal B cell activator of normal and autoantibody responses. J Immunol. 1985 Oct;135(4):2524–2533. [PubMed] [Google Scholar]
  3. Bagioni S., Sisto R., Ferraro A., Caiafa P., Turano C. A new method for the preparation of DNA--cellulose. Anal Biochem. 1978 Sep;89(2):616–619. doi: 10.1016/0003-2697(78)90390-1. [DOI] [PubMed] [Google Scholar]
  4. Bennett R. M., Gabor G. T., Merritt M. M. DNA binding to human leukocytes. Evidence for a receptor-mediated association, internalization, and degradation of DNA. J Clin Invest. 1985 Dec;76(6):2182–2190. doi: 10.1172/JCI112226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blanc B. Biochemical aspects of human milk--comparison with bovine milk. World Rev Nutr Diet. 1981;36:1–89. doi: 10.1159/000393152. [DOI] [PubMed] [Google Scholar]
  6. Bruder G., Jarasch E. D., Heid H. W. High concentrations of antibodies to xanthine oxidase in human and animal sera. Molecular characterization. J Clin Invest. 1984 Sep;74(3):783–794. doi: 10.1172/JCI111494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Concha C., Holmberg O., Morein B. Proportion of B- and T-lymphocytes in normal bovine milk. J Dairy Res. 1978 Jun;45(2):287–290. doi: 10.1017/s0022029900016472. [DOI] [PubMed] [Google Scholar]
  8. ERDOES E. G., WOHLER I. M., LEVINE I. M., WESTERMAN M. P. CARBOXYPEPTIDASE IN BLOOD AND OTHER FLUIDS. VALUES IN HUMAN BLOOD IN NORMAL AND PATHOLOGICAL CONDITIONS. Clin Chim Acta. 1965 Jan;11:39–43. doi: 10.1016/0009-8981(65)90087-2. [DOI] [PubMed] [Google Scholar]
  9. Emlen W., Mannik M. Clearance of circulating DNA-anti-DNA immune complexes in mice. J Exp Med. 1982 Apr 1;155(4):1210–1215. doi: 10.1084/jem.155.4.1210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fritzler M. J., Tan E. M. Antibodies to histones in drug-induced and idiopathic lupus erythematosus. J Clin Invest. 1978 Sep;62(3):560–567. doi: 10.1172/JCI109161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gohill J., Cary P. D., Couppez M., Fritzler M. J. Antibodies from patients with drug-induced and idiopathic lupus erythematosus react with epitopes restricted to the amino and carboxyl termini of histone. J Immunol. 1985 Nov;135(5):3116–3121. [PubMed] [Google Scholar]
  12. Hoch S. O., Longmire R. L., Hoch J. A. Unique DNA-binding protein in the serum of patients with various neoplasms. Nature. 1975 Jun 12;255(5509):560–562. doi: 10.1038/255560a0. [DOI] [PubMed] [Google Scholar]
  13. Holers V. M., Kotzin B. L. Human peripheral blood monocytes display surface antigens recognized by monoclonal antinuclear antibodies. J Clin Invest. 1985 Sep;76(3):991–998. doi: 10.1172/JCI112100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horneland M., Rekvig O. P., Jørgensen L., Hannestad K. Cultured human endothelial cells display an antigen that is recognized by certain human anti-chromatin autoantibodies. Clin Exp Immunol. 1983 Nov;54(2):373–377. [PMC free article] [PubMed] [Google Scholar]
  15. Jacob L., Tron F., Bach J. F., Louvard D. A monoclonal anti-DNA antibody also binds to cell-surface protein(s). Proc Natl Acad Sci U S A. 1984 Jun;81(12):3843–3845. doi: 10.1073/pnas.81.12.3843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jarasch E. D., Bruder G., Keenan T. W., Franke W. W. Redox constituents in milk fat globule membranes and rough endoplasmic reticulum from lactating mammary gland. J Cell Biol. 1977 Apr;73(1):223–241. doi: 10.1083/jcb.73.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kotzin B. L., Lafferty J. A., Portanova J. P., Rubin R. L., Tan E. M. Monoclonal anti-histone autoantibodies derived from murine models of lupus. J Immunol. 1984 Nov;133(5):2554–2559. [PubMed] [Google Scholar]
  18. 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]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Lewis J. G., André C. M. A serum DNA-binding protein absent in malignant disease. FEBS Lett. 1978 Aug 15;92(2):211–213. doi: 10.1016/0014-5793(78)80756-x. [DOI] [PubMed] [Google Scholar]
  21. Lönnerdal B., Keen C. L., Hurley L. S. Manganese binding proteins in human and cow's milk. Am J Clin Nutr. 1985 Mar;41(3):550–559. doi: 10.1093/ajcn/41.3.550. [DOI] [PubMed] [Google Scholar]
  22. McConahey P. J., Dixon F. J. Radioiodination of proteins by the use of the chloramine-T method. Methods Enzymol. 1980;70(A):210–213. doi: 10.1016/s0076-6879(80)70050-2. [DOI] [PubMed] [Google Scholar]
  23. McCoubrey-Hoyer A., Okarma T. B., Holman H. R. Partial purification and characterization of plasma DNA and its relation to disease activity in systemic lupus erythematosus. Am J Med. 1984 Jul;77(1):23–34. doi: 10.1016/0002-9343(84)90431-5. [DOI] [PubMed] [Google Scholar]
  24. McGhee J. D., Felsenfeld G. Nucleosome structure. Annu Rev Biochem. 1980;49:1115–1156. doi: 10.1146/annurev.bi.49.070180.005343. [DOI] [PubMed] [Google Scholar]
  25. Miskimins W. K., Roberts M. P., McClelland A., Ruddle F. H. Use of a protein-blotting procedure and a specific DNA probe to identify nuclear proteins that recognize the promoter region of the transferrin receptor gene. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6741–6744. doi: 10.1073/pnas.82.20.6741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Raptis L., Menard H. A. Quantitation and characterization of plasma DNA in normals and patients with systemic lupus erythematosus. J Clin Invest. 1980 Dec;66(6):1391–1399. doi: 10.1172/JCI109992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rekvig O. P., Hannestad K. Human autoantibodies that react with both cell nuclei and plasma membranes display specificity for the octamer of histones H2A, H2B, H3, and H4 in high salt. J Exp Med. 1980 Dec 1;152(6):1720–1733. doi: 10.1084/jem.152.6.1720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rogers J. C., Boldt D., Kornfeld S., Skinner A., Valeri C. R. Excretion of deoxyribonucleic acid by lymphocytes stimulated with phytohemagglutinin or antigen. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1685–1689. doi: 10.1073/pnas.69.7.1685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rogers J. C., Kawahara R. S. Trypsin-sensitive proteins associated with the process of unstable DNA release from human lymphocytes. Exp Cell Res. 1981 Jul;134(1):1–13. doi: 10.1016/0014-4827(81)90457-2. [DOI] [PubMed] [Google Scholar]
  30. Rubin R. L., Joslin F. G., Tan E. M. An improved ELISA for anti-native DNA by elimination of interference by anti-histone antibodies. J Immunol Methods. 1983 Oct 28;63(3):359–366. doi: 10.1016/s0022-1759(83)80009-x. [DOI] [PubMed] [Google Scholar]
  31. Rubin R. L., McNally E. M., Nusinow S. R., Robinson C. A., Tan E. M. IgG antibodies to the histone complex H2A-H2B characterize procainamide-induced lupus. Clin Immunol Immunopathol. 1985 Jul;36(1):49–59. doi: 10.1016/0090-1229(85)90038-8. [DOI] [PubMed] [Google Scholar]
  32. Sanguansermsri J., György P., Zilliken F. Polyamines in human and cow's milk. Am J Clin Nutr. 1974 Aug;27(8):859–865. doi: 10.1093/ajcn/27.8.859. [DOI] [PubMed] [Google Scholar]
  33. Schubert D., LaCorbiere M. Isolation of an adhesion-mediating protein from chick neural retina adherons. J Cell Biol. 1985 Sep;101(3):1071–1077. doi: 10.1083/jcb.101.3.1071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shahani K. M., Kwan A. J., Friend B. A. Role and significance of enzymes in human milk. Am J Clin Nutr. 1980 Aug;33(8):1861–1868. doi: 10.1093/ajcn/33.8.1861. [DOI] [PubMed] [Google Scholar]
  35. Spinola S. M., Cannon J. G. Different blocking agents cause variation in the immunologic detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 1985 Jul 16;81(1):161–165. doi: 10.1016/0022-1759(85)90132-2. [DOI] [PubMed] [Google Scholar]
  36. Tan E. M., Schur P. H., Carr R. I., Kunkel H. G. Deoxybonucleic acid (DNA) and antibodies to DNA in the serum of patients with systemic lupus erythematosus. J Clin Invest. 1966 Nov;45(11):1732–1740. doi: 10.1172/JCI105479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Thomas J. O., Wilson C. M., Hardin J. A. The major core histone antigenic determinants in systemic lupus erythematosus are in the trypsin-sensitive regions. FEBS Lett. 1984 Apr 9;169(1):90–96. doi: 10.1016/0014-5793(84)80295-1. [DOI] [PubMed] [Google Scholar]
  38. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Watson D. K., Moudrianakis E. N. Histone-dependent reconstitution and nucleosomal localization of a nonhistone chromosomal protein: the H2A-specific protease. Biochemistry. 1982 Jan 19;21(2):248–256. doi: 10.1021/bi00531a008. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES