TY - JOUR
T1 - Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy
T2 - The Raloxifene Case
AU - Mota, Cristiano
AU - Diniz, Ana
AU - Coelho, Catarina
AU - Santos-Silva, Teresa
AU - Esmaeeli, Mariam
AU - Leimkühler, Silke
AU - Cabrita, Eurico J.
AU - Marcelo, Filipa
AU - Romão, Maria João
N1 - Funding Information:
info:eu-repo/grantAgreement/EC/H2020/653706/EU#
The authors thank the Fundação para a Ciência e Tecnologia, through project PTDC/BBB-BEP/1185/2014, the IF contract to FM (IF/00780/2015), and the Ph.D. grant to AD (PD/BD/142847/2018). S.L. thanks the Deutsche Forschungsgemeinschaft (DFG) for the support with grant LE1171/8. This work is financed by National Funds from FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB.
The NMR spectrometers are part of the National NMR Network (PT NMR) and are partially supported by the Infrastructure Project No 22161 (cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC).
Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/9/9
Y1 - 2021/9/9
N2 - Human aldehyde oxidase (hAOX1) is mainly present in the liver and has an emerging role in drug metabolism, since it accepts a wide range of molecules as substrates and inhibitors. Herein, we employed an integrative approach by combining NMR, X-ray crystallography, and enzyme inhibition kinetics to understand the inhibition modes of three hAOX1 inhibitors - thioridazine, benzamidine, and raloxifene. These integrative data indicate that thioridazine is a noncompetitive inhibitor, while benzamidine presents a mixed type of inhibition. Additionally, we describe the first crystal structure of hAOX1 in complex with raloxifene. Raloxifene binds tightly at the entrance of the substrate tunnel, stabilizing the flexible entrance gates and elucidating an unusual substrate-dependent mechanism of inhibition with potential impact on drug-drug interactions. This study can be considered as a proof-of-concept for an efficient experimental screening of prospective substrates and inhibitors of hAOX1 relevant in drug discovery.
AB - Human aldehyde oxidase (hAOX1) is mainly present in the liver and has an emerging role in drug metabolism, since it accepts a wide range of molecules as substrates and inhibitors. Herein, we employed an integrative approach by combining NMR, X-ray crystallography, and enzyme inhibition kinetics to understand the inhibition modes of three hAOX1 inhibitors - thioridazine, benzamidine, and raloxifene. These integrative data indicate that thioridazine is a noncompetitive inhibitor, while benzamidine presents a mixed type of inhibition. Additionally, we describe the first crystal structure of hAOX1 in complex with raloxifene. Raloxifene binds tightly at the entrance of the substrate tunnel, stabilizing the flexible entrance gates and elucidating an unusual substrate-dependent mechanism of inhibition with potential impact on drug-drug interactions. This study can be considered as a proof-of-concept for an efficient experimental screening of prospective substrates and inhibitors of hAOX1 relevant in drug discovery.
UR - http://www.scopus.com/inward/record.url?scp=85114424221&partnerID=8YFLogxK
U2 - 10.1021/acs.jmedchem.1c01125
DO - 10.1021/acs.jmedchem.1c01125
M3 - Article
C2 - 34415167
AN - SCOPUS:85114424221
SN - 0022-2623
VL - 64
SP - 13025
EP - 13037
JO - Journal Of Medicinal Chemistry
JF - Journal Of Medicinal Chemistry
IS - 17
ER -