5'-Methylthioadenosine Nucleosidase from Yellow Lupine (Lupinus luteus): Molecular Characterization and Mutational Analysis
Authors: Bretes, Ewa; Guranowski, Andrzej; Nuc, Katarzyna
Source: Protein and Peptide Letters, Volume 18, Number 8, August 2011 , pp. 817-824(8)
Publisher: Bentham Science Publishers
Abstract:This is report of mutational analysis of higher plant 5'-methylthioadenosine nucleosidase (MTAN). We identified and characterized the gene encoding yellow lupine (Lupinus luteus) MTAN (LlMTAN). The role of active site amino acids residues Glu24, Phe134, Glu188 and Asp211 was analyzed by site-directed mutagenesis. The Glu24Gln and Asp211Asn substitutions completely abolished the enzyme activity. The Glu188Gln mutant showed only trace activity toward 5'-methylthioadenosine. These results indicate that these three amino acid residues are necessary for enzyme activity. Furthermore, as the result of replacement of Phe134 by less bulky leucine, LlMTAN acquired the ability to bind and hydrolyze S-adenosylhomocysteine. We also analyzed the sequence of the LlMTAN promoter region. It appeared that there may be a direct link between LlMTAN expression regulation and sulfate metabolism.
Keywords: 5'-Methylthioadenosine; 5'-Methylthioadenosine nucleosidase; His-Mag agarose beads; LlMTAN; Lupinus luteus; MTA cycle; Plant Cis-acting Regulatory DNA Elements; Site-directed mutagenesis; TALON resin; analogous; clones; exons; gene encoding; gene expression; lupine; sulfur-compounds; variants
Document Type: Research Article
Publication date: August 2011
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