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–