In order to analyse the amino acid determinants which alter the positional specificity of plant lipoxygenases (LOXs), multiple LOX sequence alignments and structural modelling of the enzyme-substrate interactions were carried out. These alignments suggested three amino acid residues as the primary determinants of positional specificity. Here we show the generation of two plant LOXs with new positional specificities, a Δ-linoleneate 6-LOX and an arachidonate 11-LOX, by altering only one of these determinants within the active site of two plant LOXs. In the past, site-directed-mutagenesis studies have mainly been carried out with mammalian lipoxygenases (LOXs) [1]. In these experiments two regions have been identified in the primary structure containing sequence determinants for positional specificity. Amino acids aligning with the Sloane determinants [2] are highly conserved among plant LOXs. In contrast, there is amino acid hetero-geneity among plant LOXs at the position that aligns with P353 of the rabbit reticulocyte 15-LOX (Borngräber determinants) [3].
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Conference Article|
December 01 2000
Creating lipoxygenases with new positional specificities by site-directed mutagenesis
E. Hornung;
E. Hornung
*Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany
‡Institute of Biochemistry, University Clinics Charite, Humboldt University, D-10115 Berlin, Germany
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S. Rosahl;
S. Rosahl
†Institute of Plant Biochemistry, D-06120 Halle/Saale, Germany
‡Institute of Biochemistry, University Clinics Charite, Humboldt University, D-10115 Berlin, Germany
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H. Kühn;
H. Kühn
*Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany
‡Institute of Biochemistry, University Clinics Charite, Humboldt University, D-10115 Berlin, Germany
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I. Feussner
I. Feussner
*Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany
‡Institute of Biochemistry, University Clinics Charite, Humboldt University, D-10115 Berlin, Germany
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Publisher: Portland Press Ltd
Received:
June 20 2000
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2000 Biochemical Society
2000
Biochem Soc Trans (2000) 28 (6): 825–826.
Article history
Received:
June 20 2000
Citation
E. Hornung, S. Rosahl, H. Kühn, I. Feussner; Creating lipoxygenases with new positional specificities by site-directed mutagenesis. Biochem Soc Trans 1 December 2000; 28 (6): 825–826. doi: https://doi.org/10.1042/bst0280825
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