A novel role for iron in modulating the activity and membrane-binding ability of a trimmed soybean lipoxygenase-1

Enrico Dainese, Clotilde B. Angelucci, Annalaura Sabatucci, Vincenzo De Filippis, Giampiero Mei, Mauro Maccarrone

Research output: Contribution to journalArticlepeer-review


Lipoxygenases (LOXs) are iron-containing enzymes that play critical roles in plants and animals. As yet, metal atom extraction, reconstitution, and substitution have not been successfully applied to soybean LOX-1 [Glycine max (L.) Merrill], a prototype member of the LOX family that is widely used in structural and kinetic studies. Here, tryptic digestion of native LOX-1, used as a control, allowed us to isolate the 60-kDa C-terminal region (termed miniLOX), that retains the catalytically active iron in a more accessible position. Then, iron was removed to obtain an unprecedented apo-miniLOX, which was reconstituted and substituted with different metal ions. These forms of miniLOX were characterized vs. native LOX-1 by kinetic analysis, near UV circular dichroism, steady-state fluorescence, and fluorescence resonance energy transfer. MiniLOX showed a 2-fold increase in the membrane-binding affinity compared with native LOX-1 and a remarkable 4-fold increase compared with apominiLOX (Kd=9.2±1.0, 17.9±2.0, and 45.4±4.3 μM, respectively). Furthermore, miniLOX reconstituted with Fe(II) or Fe(III) partially recovered its membrane-binding ability (Kd=21.4±2.4 and 18.9±5.5 μM, respectively), overall supporting a novel noncatalytic role for iron in the LOX family.

Original languageEnglish
Pages (from-to)1725-1736
Number of pages12
JournalFASEB Journal
Issue number6
Publication statusPublished - Jun 2010


  • Enzyme activity
  • Enzyme structure
  • Limited proteolysis
  • Metal substitution
  • Pathogen infection

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology


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