PMID- 33020066
OWN - NLM
STAT- MEDLINE
DCOM- 20210913
LR  - 20211204
IS  - 1521-009X (Electronic)
IS  - 0090-9556 (Print)
IS  - 0090-9556 (Linking)
VI  - 48
IP  - 12
DP  - 2020 Dec
TI  - Site-Directed Mutagenesis at the Molybdenum Pterin Cofactor Site of the Human 
      Aldehyde Oxidase: Interrogating the Kinetic Differences Between Human and 
      Cynomolgus Monkey.
PG  - 1364-1371
LID - 10.1124/dmd.120.000187 [doi]
AB  - The estimation of the drug clearance by aldehyde oxidase (AO) has been 
      complicated because of this enzyme's atypical kinetics and species and substrate 
      specificity. Since human AO (hAO) and cynomolgus monkey AO (mAO) have a 95.1% 
      sequence identity, cynomolgus monkeys may be the best species for estimating AO 
      clearance in humans. Here, O(6)-benzylguanine (O6BG) and dantrolene were used 
      under anaerobic conditions, as oxidative and reductive substrates of AO, 
      respectively, to compare and contrast the kinetics of these two species through 
      numerical modeling. Whereas dantrolene reduction followed the same linear 
      kinetics in both species, the oxidation rate of O6BG was also linear in mAO and 
      did not follow the already established biphasic kinetics of hAO. In an attempt to 
      determine why hAO and mAO are kinetically distinct, we have altered the hAO V811 
      and F885 amino acids at the oxidation site adjacent to the molybdenum pterin 
      cofactor to the corresponding alanine and leucine in mAO, respectively. Although 
      some shift to a more monkey-like kinetics was observed for the V811A mutant, five 
      more mutations around the AO cofactors still need to be investigated for this 
      purpose. In comparing the oxidative and reductive rates of metabolism under 
      anaerobic conditions, we have come to the conclusion that despite having similar 
      rates of reduction (4-fold difference), the oxidation rate in mAO is more than 
      50-fold slower than hAO. This finding implies that the presence of nonlinearity 
      in AO kinetics is dependent upon the degree of imbalance between the rates of 
      oxidation and reduction in this enzyme. SIGNIFICANCE STATEMENT: Although they 
      have as much as 95.1% sequence identity, human and cynomolgus monkey aldehyde 
      oxidase are kinetically distinct. Therefore, monkeys may not be good estimators 
      of drug clearance in humans.
CI  - Copyright (c) 2020 by The American Society for Pharmacology and Experimental 
      Therapeutics.
FAU - Abbasi, Armina
AU  - Abbasi A
AD  - Department of Chemistry, Washington State University, Pullman, Washington.
FAU - Joswig-Jones, Carolyn A
AU  - Joswig-Jones CA
AD  - Department of Chemistry, Washington State University, Pullman, Washington.
FAU - Jones, Jeffrey P
AU  - Jones JP
AD  - Department of Chemistry, Washington State University, Pullman, Washington 
      jpj@wsu.edu.
LA  - eng
GR  - R01 GM100874/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20201005
PL  - United States
TA  - Drug Metab Dispos
JT  - Drug metabolism and disposition: the biological fate of chemicals
JID - 9421550
RN  - 0 (Coenzymes)
RN  - 0 (Metalloproteins)
RN  - 0 (Molybdenum Cofactors)
RN  - 0 (Pteridines)
RN  - 01KC87F8FE (O(6)-benzylguanine)
RN  - 5Z93L87A1R (Guanine)
RN  - ATN6EG42UQ (molybdenum cofactor)
RN  - EC 1.2.3.1 (AOX1 protein, human)
RN  - EC 1.2.3.1 (Aldehyde Oxidase)
RN  - F64QU97QCR (Dantrolene)
SB  - IM
MH  - Aldehyde Oxidase/genetics/*metabolism
MH  - Animals
MH  - Coenzymes/*metabolism
MH  - Dantrolene/pharmacokinetics
MH  - Drug Evaluation, Preclinical/methods
MH  - Guanine/analogs & derivatives/pharmacokinetics
MH  - Macaca fascicularis/genetics
MH  - Metalloproteins/*metabolism
MH  - Molybdenum Cofactors
MH  - Mutagenesis, Site-Directed
MH  - Oxidation-Reduction
MH  - Pteridines/*metabolism
MH  - Sequence Homology, Amino Acid
MH  - Species Specificity
MH  - Substrate Specificity/genetics
PMC - PMC7718725
EDAT- 2020/10/07 06:00
MHDA- 2021/09/14 06:00
PMCR- 2021/12/01
CRDT- 2020/10/06 05:25
PHST- 2020/07/17 00:00 [received]
PHST- 2020/09/25 00:00 [accepted]
PHST- 2020/10/07 06:00 [pubmed]
PHST- 2021/09/14 06:00 [medline]
PHST- 2020/10/06 05:25 [entrez]
PHST- 2021/12/01 00:00 [pmc-release]
AID - dmd.120.000187 [pii]
AID - DMD_AR2020000187 [pii]
AID - 10.1124/dmd.120.000187 [doi]
PST - ppublish
SO  - Drug Metab Dispos. 2020 Dec;48(12):1364-1371. doi: 10.1124/dmd.120.000187. Epub 
      2020 Oct 5.