Identification of the iron-binding histidine residues in soybean lipoxygenase L-1.
Lipoxygenases constitute a class of non-heme, non-sulfur iron dioxygenases acting upon lipids possessing a 1,4-cis-cis-pentadiene moiety. The iron is known to be essential for activity. A motif of six histidine residues has been found in all of the thirteen lipoxygenases, from both plant and animal sources, whose sequences have been reported. We had previously obtained mutant proteins in which each of the 6 conserved histidines of soybean lipoxygenase L-1 had been replaced and found that the mutants H499Q, H504Q (or H504S) and H690Q had no detectable enzymatic activity. We have now found that these inactive proteins contain no Fe, although they have the same electrophoretic mobility as wild-type L-1 under both denaturing and non-denaturing conditions and react with anti-L-1 antibodies.
Steczko
J
J
Department of Biochemistry, Purdue University West Lafayette, Indiana 47906-1153.
Axelrod
B
B
D000595:Amino Acid Sequence; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine; D007501:Iron / Q000378:metabolism*; D008084:Lipoxygenase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013025:Glycine max / Q000201:enzymology* / Q000235:genetics
D011994:Recombinant Proteins; D006639:Histidine; D007501:Iron; D008084:Lipoxygenase