title: Dimerization of human 5-lipoxygenase.
abstract: Human 5-lipoxygenase (5-LO) can form dimers as shown here via native gel electrophoresis, gel filtration chromatography and LILBID (laser induced liquid bead ion desorption) mass spectrometry. After glutathionylation of 5-LO by diamide/glutathione treatment, dimeric 5-LO was no longer detectable and 5-LO almost exclusively exists in the monomeric form which showed full catalytic activity. Incubation of 5-LO with diamide alone led to a disulfide-bridged dimer and to oligomer formation which displays a strongly reduced catalytic activity. The bioinformatic analysis of the 5-LO surface for putative protein-protein interaction domains and molecular modeling of the dimer interface suggests a head to tail orientation of the dimer which also explains the localization of previously reported ATP binding sites. This interface domain was confirmed by the observation that 5-LO dimer formation and inhibition of activity by diamide was largely prevented when four cysteines (C159S, C300S, C416S, C418S) in this domain were mutated to serines.
authors: Institute of Pharmaceutical Chemistry/ZAFES, University of Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany.
authors: Ann-Kathrin
authors: AK
authors: Häfner
authors: Mihaela
authors: M
authors: Cernescu
authors: Bettina
authors: B
authors: Hofmann
authors: Michael
authors: M
authors: Ermisch
authors: Michael
authors: M
authors: Hörnig
authors: Julia
authors: J
authors: Metzner
authors: Gisbert
authors: G
authors: Schneider
authors: Bernhard
authors: B
authors: Brutschy
authors: Dieter
authors: D
authors: Steinhilber
mesh_terms: D001094:Arachidonate 5-Lipoxygenase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D003958:Diamide / Q000737:chemistry; D019075:Electrophoresis, Capillary; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry; D008958:Models, Molecular; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D003958:Diamide; D001094:Arachidonate 5-Lipoxygenase; D005978:Glutathione