Dimerization of human 5-lipoxygenase

AK Häfner, M Cernescu, B Hofmann, M Ermisch… - 2011 - degruyter.com
AK Häfner, M Cernescu, B Hofmann, M Ermisch, M Hörnig, J Metzner, G Schneider
2011degruyter.com
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 …
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.
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