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. 1994 Jan 18;91(2):689–693. doi: 10.1073/pnas.91.2.689

Photoaffinity labeling of Arabidopsis thaliana plasma membrane vesicles by 5-azido-[7-3H]indole-3-acetic acid: identification of a glutathione S-transferase.

R Zettl 1, J Schell 1, K Palme 1
PMCID: PMC43014  PMID: 8290582

Abstract

We used 5-azido-[7-3H]indole-3-acetic acid (5-azido-[7-3H]IAA), a photoaffinity analogue of the plant hormone indole-3-acetic acid (IAA), to search for auxin-binding proteins in Arabidopsis thaliana membranes. We identified an auxin-binding protein with a molecular mass of 24 kDa (Atpm24) in microsomes as well as in plasma membrane vesicles. Atpm24 was solubilized by 1% Triton X-100 and partially purified. A cDNA clone (Atpm24.1) corresponding to Atpm24 was isolated. The amino acid sequence predicted from the Atpm24.1 cDNA contains 212 amino acid residues with a relative molecular mass of 24,128 Da. Data base searches revealed that the predicted protein has homology to glutathione S-transferases (GSTs; EC 2.5.1.18). When Atpm24.1 was expressed in Escherichia coli, we found a high level of GST activity in the bacterial extracts. We have analyzed the substrate specificity of this protein and found that cumene hydroperoxide and trans-stilbene oxide but not trans-cinnamic acid or IAA-CoA were substrates. A role for this GST in physiological processes of plants is discussed.

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Selected References

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  1. Bhargava M. M., Listowsky I., Arias I. M. Studies on subunit structure and evidence that ligandin is a heterodimer. J Biol Chem. 1978 Jun 25;253(12):4116–4119. [PubMed] [Google Scholar]
  2. Bilang J., Macdonald H., King P. J., Sturm A. A soluble auxin-binding protein from Hyoscyamus muticus is a glutathione S-transferase. Plant Physiol. 1993 May;102(1):29–34. doi: 10.1104/pp.102.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DeJong J. L., Morgenstern R., Jörnvall H., DePierre J. W., Tu C. P. Gene expression of rat and human microsomal glutathione S-transferases. J Biol Chem. 1988 Jun 15;263(17):8430–8436. [PubMed] [Google Scholar]
  4. Dudler R., Hertig C., Rebmann G., Bull J., Mauch F. A pathogen-induced wheat gene encodes a protein homologous to glutathione-S-transferases. Mol Plant Microbe Interact. 1991 Jan-Feb;4(1):14–18. doi: 10.1094/mpmi-4-014. [DOI] [PubMed] [Google Scholar]
  5. Feldwisch J., Zettl R., Hesse F., Schell J., Palme K. An auxin-binding protein is localized to the plasma membrane of maize coleoptile cells: identification by photoaffinity labeling and purification of a 23-kda polypeptide. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):475–479. doi: 10.1073/pnas.89.2.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grove G., Zarlengo R. P., Timmerman K. P., Li N. Q., Tam M. F., Tu C. P. Characterization and heterospecific expression of cDNA clones of genes in the maize GSH S-transferase multigene family. Nucleic Acids Res. 1988 Jan 25;16(2):425–438. doi: 10.1093/nar/16.2.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hicks G. R., Rayle D. L., Jones A. M., Lomax T. L. Specific photoaffinity labeling of two plasma membrane polypeptides with an azido auxin. Proc Natl Acad Sci U S A. 1989 Jul;86:4948–4952. doi: 10.1073/pnas.86.13.4948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hicks G. R., Rayle D. L., Lomax T. L. The diageotropica mutant of tomato lacks high specific activity auxin binding sites. Science. 1989 Jul 7;245:52–54. doi: 10.1126/science.245.4913.52. [DOI] [PubMed] [Google Scholar]
  9. Jones A. M., Venis M. A. Photoaffinity labeling of indole-3-acetic acid-binding proteins in maize. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6153–6156. doi: 10.1073/pnas.86.16.6153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Macdonald H., Jones A. M., King P. J. Photoaffinity labeling of soluble auxin-binding proteins. J Biol Chem. 1991 Apr 25;266(12):7393–7399. [PubMed] [Google Scholar]
  11. Mannervik B., Danielson U. H. Glutathione transferases--structure and catalytic activity. CRC Crit Rev Biochem. 1988;23(3):283–337. doi: 10.3109/10409238809088226. [DOI] [PubMed] [Google Scholar]
  12. Morgenstern R., DePierre J. W. Microsomal glutathione transferase. Purification in unactivated form and further characterization of the activation process, substrate specificity and amino acid composition. Eur J Biochem. 1983 Aug 15;134(3):591–597. doi: 10.1111/j.1432-1033.1983.tb07607.x. [DOI] [PubMed] [Google Scholar]
  13. Morgenstern R., Guthenberg C., Depierre J. W. Microsomal glutathione S-transferase. Purification, initial characterization and demonstration that it is not identical to the cytosolic glutathione S-transferases A, B and C. Eur J Biochem. 1982 Nov;128(1):243–248. [PubMed] [Google Scholar]
  14. Morgenstern R., Meijer J., Depierre J. W., Ernster L. Characterization of rat-liver microsomal glutathione S-transferase activity. Eur J Biochem. 1980 Feb;104(1):167–174. doi: 10.1111/j.1432-1033.1980.tb04412.x. [DOI] [PubMed] [Google Scholar]
  15. Reinemer P., Dirr H. W., Ladenstein R., Huber R., Lo Bello M., Federici G., Parker M. W. Three-dimensional structure of class pi glutathione S-transferase from human placenta in complex with S-hexylglutathione at 2.8 A resolution. J Mol Biol. 1992 Sep 5;227(1):214–226. doi: 10.1016/0022-2836(92)90692-d. [DOI] [PubMed] [Google Scholar]
  16. Reinemer P., Dirr H. W., Ladenstein R., Schäffer J., Gallay O., Huber R. The three-dimensional structure of class pi glutathione S-transferase in complex with glutathione sulfonate at 2.3 A resolution. EMBO J. 1991 Aug;10(8):1997–2005. doi: 10.1002/j.1460-2075.1991.tb07729.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Samuelsson B. Leukotrienes: mediators of immediate hypersensitivity reactions and inflammation. Science. 1983 May 6;220(4597):568–575. doi: 10.1126/science.6301011. [DOI] [PubMed] [Google Scholar]
  18. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  19. Takahashi Y., Nagata T. parB: an auxin-regulated gene encoding glutathione S-transferase. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):56–59. doi: 10.1073/pnas.89.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zhou J., Goldsbrough P. B. An Arabidopsis gene with homology to glutathione S-transferases is regulated by ethylene. Plant Mol Biol. 1993 Jun;22(3):517–523. doi: 10.1007/BF00015980. [DOI] [PubMed] [Google Scholar]

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