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Inhibitory effects of auxins and related substances on the activity of an Arabidopsis glutathione S‐transferase isozyme expressed in Escherichia coli
Author(s) -
Watahiki Masaaki K.,
Mori Hitoshi,
Yamamoto Kotaro T.
Publication year - 1995
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1995.tb00969.x
Subject(s) - glutathione , auxin , biochemistry , escherichia coli , isozyme , glutathione s transferase , arabidopsis , enzyme , biology , 2,4 dichlorophenoxyacetic acid , complementary dna , microbiology and biotechnology , chemistry , gene , mutant , botany
Glutathione S‐transferases (GSTs; EC 2.5.1.18) are encoded by a gene family. Some GSTs have the capacity to bind to indole‐3‐acetic acid (IAA), whereas the gene expression of other GSTs is regulated by auxin. In order to assess a possible physiological significance of the auxin binding of GST, we investigated effects of auxins on the activity of GST expressed in Escherichia coli. cDNA cloning was carried out for the fifth gene ( GST5 ) of GST in Arabidopsis. Although the deduced amino acid sequence of GST5 was remotely related to that of the other Arabidopsis GSTs (less than 20% identical), the GST5 protein (GST5) expressed in E. coli showed GST activity. Apparent K m values of GST5 are 0.86 and 1.29 m M for glutathione (GSH) and 1‐chloro‐2,4‐dinitrobenzene, respectively. IAA, 2,4‐dichlorophenoxyacetic acid (2,4‐D), 1‐naphthaleneacetic acid (1‐NAA) and 2‐NAA inhibited the enzyme activity competitively with respect to GSH. The apparent K i of IAA is 1.56 m M . Salicylic acid inhibited GST activity in a noncompetitive manner. 2,4‐D was the most inhibitory among the tested chemicals. GST5 bound to GSH‐immobilized agarose gel was effectively eluted by IAA. These results indicate that IAA and the related substances bind to GST5 at the GSH‐binding site, and exclude the possibility that the compounds could be substrates for GST5. Although the K i value of IAA is too high for any physiological consequences, it might be assumed that GST activity is modulated in vivo by an auxin‐related substance(s). The steady‐state level of the GST5 mRNA was increased by wounding, heat shock, and spraying buffer on the plant, but was not influenced by auxin treatment.