Isolation of lipoxygenase isoforms from Glycine max embryo axes based on apparent cross-reactivity with anti-myosin antibodies.
Three lipoxygenase isoforms were isolated from Glycine max embryo axes. A number of proteins around 97 kDa cross-reacted with several anti-actin and anti-myosin antibodies and these were used to follow their purification through gel filtration, hydroxyapatite and anion exchange columns. The 97-kDa cross-reactive material eluted in the unbound fractions of the last anion exchange column, and displayed two components of pI's 6.2 and 6.3. Further phase partition of this fraction in TX-114 yielded a hydrophobic 97 kDa protein. Additionally, a 95-kDa protein was retained and eluted from this last column. Partial peptide sequences indicated that the 95 kDa protein was soybean lipoxygenase-1, the first 97 kDa protein was lypoxygenase-3, and the hydrophobic 97 kDa protein was lipoxygenase-2. Several possible reasons for the cross-reactivity with the antibodies are discussed. To our knowledge, this is the first example of individual lipoxygenase isoforms isolated from soybean embryo axes.
Islas-Flores
Ignacio
I
Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, U.N.A.M., Cuernavaca, Mexico.
Corrales-Villamar
Salvador
S
Bearer
Elaine
E
Raya
Juan Carlos
JC
Villanueva
Marco A
MA
D000199:Actins / Q000276:immunology; D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western; D003429:Cross Reactions; D018525:Germination; D007527:Isoenzymes / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D008084:Lipoxygenase / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009218:Myosins / Q000276:immunology; D012639:Seeds / Q000201:enzymology; D013025:Glycine max / Q000196:embryology / Q000201:enzymology*
D000199:Actins; D000906:Antibodies; D007527:Isoenzymes; D008084:Lipoxygenase; D009218:Myosins