title: Regio- and stereospecificity in the oxygenation of arachidonic acid catalyzed by Leu597 mutants of rabbit 15-lipoxygenase: a QM/MM study.
abstract: We combined quantum mechanics/molecular mechanics calculations with molecular dynamics simulations to study the addition of O2 to the pentadienyl radical of arachidonic acid (AA) catalyzed by the Leu597Val and Leu597Ala mutants of rabbit 15-lipoxygenase (15-rLO). In the Leu597Val mutant, the addition of O2 to C15 of AA is the predominant path, although it reduces the C15/C11 product ratio by almost ten times with respect to the wildtype enzyme. The S stereochemistry is kept. Mutation to Ala causes just the opposite effect: regiospecificity favoring addition to C15 is somewhat sharper than that in the wildtype, but the stereochemistry is R. This is because the extra space created by the mutation to Ala is big enough for AA to move so that it can adopt an alternative binding mode, and this opens new feasible paths for the attack of O2 . So, we showed that the Leu597Ala mutant of 15r-LO works as an aspirin-acetylated cyclooxygenase-2, which makes 15-(R)- hydroperoxyeicosatetraenoic acid.
authors.affiliation: Departament de Química, Universitat Autònoma de Barcelona, Bellaterra 08193, Barcelona (Spain); Institut de Biotecnologia i de Biomedicina (IBB), Universitat Autònoma de Barcelona, Bellaterra 08193, Barcelona (Spain).
authors.forename: Reynier
authors.initials: R
authors.lastname: Suardíaz
authors.forename: Laura
authors.initials: L
authors.lastname: Masgrau
authors.forename: José M
authors.initials: JM
authors.lastname: Lluch
authors.forename: Angels
authors.initials: A
authors.lastname: González-Lafont
mesh_terms: D000818:Animals; D001093:Arachidonate 15-Lipoxygenase / Q000737:chemistry* / Q000235:genetics*; D016718:Arachidonic Acid / Q000737:chemistry*; D002384:Catalysis; D051546:Cyclooxygenase 2 / Q000737:chemistry; D007930:Leucine / Q000235:genetics*; D056004:Molecular Dynamics Simulation; D010100:Oxygen / Q000737:chemistry; D011789:Quantum Theory; D011817:Rabbits; D013237:Stereoisomerism
chemical_list: D016718:Arachidonic Acid; D001093:Arachidonate 15-Lipoxygenase; D051546:Cyclooxygenase 2; D007930:Leucine; D010100:Oxygen
keywords: enzymes; mutants; peroxidation; regioselectivity; stereospecificity