Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1985 Aug 26;13(16):5833–5842. doi: 10.1093/nar/13.16.5833

Purification of antibodies for the cytokinin N6-(delta 2-isopentenyl) adenosine by affinity chromatography.

P Senapathy, C Jayabaskaran, M T Jacob
PMCID: PMC321915  PMID: 3929232

Abstract

Isopentenyl adenosine antibodies useful in the investigations of the "cytokinin" functions of isopentenyl adenosine were purified by affinity chromatography. Using different affinity columns, the antibodies were purified to near complete purity. Analyses of the purified proteins revealed the presence of isopentenyl adenosine binding proteins in normal rabbit serum, which presence supports a suggested role for isopentenyl adenosine and its related compounds in animal cell division in vivo.

Full text

PDF
5833

Images in this article

5838Image
on p.5838
5839Image
on p.5839

Selected References

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

  1. Bauvois B., Leng M. Heterogeneity of antibodies to adenosine 5'-monophosphate. Biochim Biophys Acta. 1976 Nov 12;454(1):1–8. doi: 10.1016/0005-2787(76)90349-x. [DOI] [PubMed] [Google Scholar]
  2. Drocourt J. L., Leng M. Antibodies to adenosine 5'-monophosphate: purification and specificity. Eur J Biochem. 1975 Aug 1;56(1):149–155. doi: 10.1111/j.1432-1033.1975.tb02217.x. [DOI] [PubMed] [Google Scholar]
  3. ERLANGER B. F., BEISER S. M. ANTIBODIES SPECIFIC FOR RIBONUCLEOSIDES AND RIBONUCLEOTIDES AND THEIR REACTION WITH DNA. Proc Natl Acad Sci U S A. 1964 Jul;52:68–74. doi: 10.1073/pnas.52.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gallo R. C., Hecht S. M., Whang-Peng J., O'Hopp S. N 6 -( 2 -Isopentenyl)adenosine: the regulatory effects of a cytokinin and modified nucleoside from tRNA on human lymphocytes. Biochim Biophys Acta. 1972 Nov 9;281(4):488–500. doi: 10.1016/0005-2787(72)90149-9. [DOI] [PubMed] [Google Scholar]
  5. Gallo R. C., Whang-Peng J., Perry S. Isopentenyladenosine stimulates and inhibits mitosis of human lymphocytes treated with phytohemagglutinin. Science. 1969 Jul 25;165(3891):400–402. doi: 10.1126/science.165.3891.400. [DOI] [PubMed] [Google Scholar]
  6. Hacker B., Feldbush T. L. N6-(delta-2-isopentenyl) adenosine--effects upon nucleic acid synthesis in lymphocytes in vitro and the development of immunologic hypersensitivity in vivo. Biochem Pharmacol. 1969 Apr;18(4):847–853. doi: 10.1016/0006-2952(69)90055-0. [DOI] [PubMed] [Google Scholar]
  7. Hacker B., Van Vunakis H., Levine L. Formation of an antibody with serologic specificity for N 6 -( 2 -isopentenyl) adenosine. J Immunol. 1972 Jun;108(6):1726–1728. [PubMed] [Google Scholar]
  8. Halloran M. J., Parker C. W. The preparation of nucleotide-protein conjugates: carbodiimides as coupling agents. J Immunol. 1966 Mar;96(3):373–378. [PubMed] [Google Scholar]
  9. Humanyun M. Z., Jacob T. M. Evidence for an unusual immunological response to isopentenyladenosine in rabbits. FEBS Lett. 1974 Jul 15;43(2):195–198. doi: 10.1016/0014-5793(74)80998-1. [DOI] [PubMed] [Google Scholar]
  10. Humayun M. Z., Jacob T. M. Immunologic studies on nucleic acids and their components. I. An analysis of the specificity of anti-deoxyadenylate antibodies by a membrane-binding technique. Biochim Biophys Acta. 1973 Nov 26;331(1):41–53. doi: 10.1016/0005-2787(73)90417-6. [DOI] [PubMed] [Google Scholar]
  11. Humayun M. Z., Jacob T. M. Immunologic studies on nucleic acids and their components. II. Reversible inhibition of anti-nucleoside antibodies in aqueous pyridine and its application in antibody purification. Biochim Biophys Acta. 1974 Apr 27;349(1):84–95. doi: 10.1016/0005-2787(74)90011-2. [DOI] [PubMed] [Google Scholar]
  12. Khan S. A., Humayun M. Z., Jacob T. M. A sensitive radioimmunoassay for isopentenyladenosine. Anal Biochem. 1977 Dec;83(2):632–635. doi: 10.1016/0003-2697(77)90067-7. [DOI] [PubMed] [Google Scholar]
  13. Lavayre J., Leng M. Purification and specificity of antibodies to adenosine. Biochimie. 1977;59(1):33–42. doi: 10.1016/s0300-9084(77)80083-7. [DOI] [PubMed] [Google Scholar]
  14. Lubit B. W., Pham T. D., Miller O. J., Erlanger B. F. Localization of 5-methylcytosine in human metaphase chromosomes by immunoelectron microscopy. Cell. 1976 Dec;9(4 Pt 1):503–509. doi: 10.1016/0092-8674(76)90032-5. [DOI] [PubMed] [Google Scholar]
  15. Miller O. J., Schnedl W., Allen J., Erlanger B. F. 5-Methylcytosine localised in mammalian constitutive heterochromatin. Nature. 1974 Oct 18;251(5476):636–637. doi: 10.1038/251636a0. [DOI] [PubMed] [Google Scholar]
  16. Milstone D. S., Vold B. S., Glitz D. G., Shutt N. Antibodies to N6-(delta2-isopentenyl) adenosine and its nucleotide: interaction with purified tRNAs and with bases, nucleosides and nucleotides of the isopentenyladenosine family. Nucleic Acids Res. 1978 Sep;5(9):3439–3455. doi: 10.1093/nar/5.9.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Munns T. W., Sims H. F., Liszewski M. K. Immunospecific retention of oligonucleotides possessing N6-methyladenosine and 7-methylguanosine. J Biol Chem. 1977 May 10;252(9):3102–3104. [PubMed] [Google Scholar]
  18. Olson H. M., Glitz D. G. Ribosome structure: localization of 3' end of RNA in small subunit by immunoelectronmicroscopy. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3769–3773. doi: 10.1073/pnas.76.8.3769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Olson H. M., Grant P. G., Glitz D. G., Cooperman B. S. Immunoelectron microscopic localization of the site of photo-induced affinity labeling of the small ribosomal subunit with puromycin. Proc Natl Acad Sci U S A. 1980 Feb;77(2):890–894. doi: 10.1073/pnas.77.2.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Politz S. M., Glitz D. G. Magnesium-dependent interaction of 30S ribosomal subunits with antibodies to N6, N6-dimethyladenosine. Biochemistry. 1980 Aug 5;19(16):3786–3791. doi: 10.1021/bi00557a021. [DOI] [PubMed] [Google Scholar]
  21. Politz S. M., Glitz D. G. Ribosome structure: localization of N6,N6-dimethyladenosine by electron microscopy of a ribosome-antibody complex. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1468–1472. doi: 10.1073/pnas.74.4.1468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rudkin G. T., Stollar B. D. High resolution detection of DNA-RNA hybrids in situ by indirect immunofluorescence. Nature. 1977 Feb 3;265(5593):472–473. doi: 10.1038/265472a0. [DOI] [PubMed] [Google Scholar]
  23. Schreck R. R., Erlanger B. F., Miller O. J. The use of antinucleoside antibodies to probe the organization of chromosomes denatured by ultraviolet irradiation. Exp Cell Res. 1974 Sep;88(1):31–39. doi: 10.1016/0014-4827(74)90614-4. [DOI] [PubMed] [Google Scholar]
  24. Senapathy P., Jacob M. T. Identification and purification of tRNAs containing N6-(delta 2-isopentenyl) adenosine using antibodies specific for N6-(delta-isopentenyl) adenosine. J Biol Chem. 1981 Nov 25;256(22):11580–11584. [PubMed] [Google Scholar]
  25. Suk D., Simpson C. L., Mihich E. Toxicological and antiproliferative effects of N6-(delta2-isopentenyl) adenosine, a natural component of mammalian transfer RNA. Cancer Res. 1970 May;30(5):1429–1436. [PubMed] [Google Scholar]
  26. Tsoupras G., Luu B., Hoffmann J. A. A Cytokinin (Isopentenyl-Adenosyl-Mononucleotide) Linked to Ecdysone in Newly Laid Eggs of Locusta migratoria. Science. 1983 Apr 29;220(4596):507–509. doi: 10.1126/science.220.4596.507. [DOI] [PubMed] [Google Scholar]
  27. Vold B. S., Nolen H. W., 3rd A unique method utilizing antinucleotide antibodies for evaluating changes in the levels of modified nucleosides of tRNAs from crude extracts of whole cells. Nucleic Acids Res. 1979 Oct 25;7(4):971–980. doi: 10.1093/nar/7.4.971. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES