Conformational Heterogeneity and Cooperative Effects of Mammalian ALOX15

Igor Ivanov; Alejandro Cruz; Alexander Zhuravlev; Almerinda Di Venere; Eleonora Nicolai; Sabine Stehling; José M. Lluch; Àngels González-Lafont; Hartmut Kuhn

Conformational Heterogeneity and Cooperative Effects of Mammalian ALOX15

Igor Ivanov, Alejandro Cruz, Alexander Zhuravlev, Almerinda Di Venere, Eleonora Nicolai, Sabine Stehling, José M. Lluch, Àngels González-Lafont, Hartmut Kuhn

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Abstract

Arachidonic acid lipoxygenases (ALOXs) have been suggested to function as monomeric enzymes, but more recent data on rabbit ALOX15 indicated that there is a dynamic monomer‐dimer equilibrium in aqueous solution. In the presence of an active site ligand (the ALOX15 inhibitor RS7) rabbit ALOX15 was crystalized as heterodimer and the X‐ray coordinates of the two monomers within the dimer exhibit subtle structural differences. Using native polyacrylamide electrophoresis, we here observed that highly purified and predominantly monomeric rabbit ALOX15 and human ALOX15B are present in two conformers with distinct electrophoretic mobilities. In silico docking studies, molecular dynamics simulations, site directed mutagenesis experiments and kinetic meas-urements suggested that in aqueous solutions the two enzymes exhibit motional flexibility, which may impact the enzymatic properties.

Original language	English
Article number	3285
Number of pages	18
Journal	International journal of molecular sciences
Volume	22
Issue number	6
Publication status	Published - 23 Mar 2021

Keywords

Amino Acid Substitution
Animals
Arachidonate 15-Lipoxygenase/chemistry
Catalysis
Cooperative effects
Crystal structure
Humans
Isoenzymes
Kinetics
Lipoxygenases
Models, Molecular
Molecular dynamics
Mutation
Protein Binding
Protein Conformation
Protein Interaction Domains and Motifs
Protein Multimerization
Protein–protein interactions
Rabbits
cooperative effects
crystal structure
lipoxygenases
molecular dynamics
protein interactions
protein&#8211

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title = "Conformational Heterogeneity and Cooperative Effects of Mammalian ALOX15",

abstract = "Arachidonic acid lipoxygenases (ALOXs) have been suggested to function as monomeric enzymes, but more recent data on rabbit ALOX15 indicated that there is a dynamic monomer‐dimer equilibrium in aqueous solution. In the presence of an active site ligand (the ALOX15 inhibitor RS7) rabbit ALOX15 was crystalized as heterodimer and the X‐ray coordinates of the two monomers within the dimer exhibit subtle structural differences. Using native polyacrylamide electrophoresis, we here observed that highly purified and predominantly monomeric rabbit ALOX15 and human ALOX15B are present in two conformers with distinct electrophoretic mobilities. In silico docking studies, molecular dynamics simulations, site directed mutagenesis experiments and kinetic meas-urements suggested that in aqueous solutions the two enzymes exhibit motional flexibility, which may impact the enzymatic properties.",

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author = "Igor Ivanov and Alejandro Cruz and Alexander Zhuravlev and Venere, {Almerinda Di} and Eleonora Nicolai and Sabine Stehling and Lluch, {Jos{\'e} M.} and {\`A}ngels Gonz{\'a}lez-Lafont and Hartmut Kuhn",

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AU - Ivanov, Igor

AU - Cruz, Alejandro

AU - Zhuravlev, Alexander

AU - Venere, Almerinda Di

AU - Nicolai, Eleonora

AU - Stehling, Sabine

AU - Lluch, José M.

AU - González-Lafont, Àngels

AU - Kuhn, Hartmut

PY - 2021/3/23

Y1 - 2021/3/23

N2 - Arachidonic acid lipoxygenases (ALOXs) have been suggested to function as monomeric enzymes, but more recent data on rabbit ALOX15 indicated that there is a dynamic monomer‐dimer equilibrium in aqueous solution. In the presence of an active site ligand (the ALOX15 inhibitor RS7) rabbit ALOX15 was crystalized as heterodimer and the X‐ray coordinates of the two monomers within the dimer exhibit subtle structural differences. Using native polyacrylamide electrophoresis, we here observed that highly purified and predominantly monomeric rabbit ALOX15 and human ALOX15B are present in two conformers with distinct electrophoretic mobilities. In silico docking studies, molecular dynamics simulations, site directed mutagenesis experiments and kinetic meas-urements suggested that in aqueous solutions the two enzymes exhibit motional flexibility, which may impact the enzymatic properties.

AB - Arachidonic acid lipoxygenases (ALOXs) have been suggested to function as monomeric enzymes, but more recent data on rabbit ALOX15 indicated that there is a dynamic monomer‐dimer equilibrium in aqueous solution. In the presence of an active site ligand (the ALOX15 inhibitor RS7) rabbit ALOX15 was crystalized as heterodimer and the X‐ray coordinates of the two monomers within the dimer exhibit subtle structural differences. Using native polyacrylamide electrophoresis, we here observed that highly purified and predominantly monomeric rabbit ALOX15 and human ALOX15B are present in two conformers with distinct electrophoretic mobilities. In silico docking studies, molecular dynamics simulations, site directed mutagenesis experiments and kinetic meas-urements suggested that in aqueous solutions the two enzymes exhibit motional flexibility, which may impact the enzymatic properties.

KW - Amino Acid Substitution

KW - Animals

KW - Arachidonate 15-Lipoxygenase/chemistry

KW - Catalysis

KW - Cooperative effects

KW - Crystal structure

KW - Humans

KW - Isoenzymes

KW - Kinetics

KW - Lipoxygenases

KW - Models, Molecular

KW - Molecular dynamics

KW - Mutation

KW - Protein Binding

KW - Protein Conformation

KW - Protein Interaction Domains and Motifs

KW - Protein Multimerization

KW - Protein–protein interactions

KW - Rabbits

KW - cooperative effects

KW - crystal structure

KW - lipoxygenases

KW - molecular dynamics

KW - protein interactions

KW - protein&#8211

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C2 - 33807076

SN - 1661-6596

VL - 22

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

IS - 6

M1 - 3285

ER -

Conformational Heterogeneity and Cooperative Effects of Mammalian ALOX15

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