Evidence map›Paper›PMID 40454940›Full record

ArticleThe Biochemical journal2025

A new naphthalene-based fluorogenic substrate for cytochrome P450 4A11.

Dmitri R Davydov, Kannapiran Ponraj, Nadezhda Davydova, Dilip Kumar Singh, Bhagwat Prasad

Abstract read
In one paragraph

Article in The Biochemical journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Article
  2. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Dmitri R DavydovDepartment of Chemistry, Washington State University, Pullman, WA 99164, U.S.A.ORCID 0000-0001-8520-0033
Kannapiran PonrajDepartment of Chemistry, Washington State University, Pullman, WA 99164, U.S.A.ORCID 0000-0002-7195-5432
Nadezhda DavydovaDepartment of Chemistry, Washington State University, Pullman, WA 99164, U.S.A.
Dilip Kumar SinghDepartment of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, U.S.A.ORCID 0000-0001-7248-6714
Bhagwat PrasadDepartment of Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, U.S.A.ORCID 0000-0002-9090-0912

Funding

Inter-Enzyme Crosstalk in the Cytochrome P450 Ensemble: Implications for the Effects of Alcohol on Drug Metabolism and Alcohol-Drug InteractionsR01AA030155 · NIAAA · WASHINGTON STATE UNIVERSITY · PI DAVYDOV, DMITRI R, PRASAD, BHAGWAT · 2022 to 2025
$2.0M
NIAAA NIH HHS R01 AA030155NIH HHS R01AA030155
6 · The paper itself

Abstract

We aimed to create a high-throughput fluorimetric assay for the activity of CYP4A11, the major 20-HETE-producing enzyme. To this end, we probed 3-(6-methoxynaphthalen-2-yl)acrylic acid (MONACRA) as a potential CYP4A11 substrate. We studied its metabolism using human liver microsomes (HLM) and recombinant P450 enzymes. O-demethylation of MONACRA by cytochromes P450 creates 3-(6-hydroxynaphthalen-2-yl)acrylic acid. The bright fluorescence of the product and its clear spectral resolution from the substrate allowed us to create a fluorimetric assay of MONACRA metabolism. We tested 16 recombinant human P450 enzymes and found noticeable demethylation activity only with CYP4A11 and CYP1A2. The KM for CYP4A11 is 189±37 μM, and the kcat accounts for 67±18 min-1. CYP1A2 exhibits a KM of 161±34 μM, with a kcat value of 44±6 min-1, although this enzyme also exhibited a decreased rate of turnover at high substrate concentrations, evidencing substrate inhibition with Ksi=650±200 μM. The studies with fluvoxamine and epalrestat, specific inhibitors of CYP1A2 and CYP4A11, respectively, showed that despite the activity of recombinant CYP1A2 with MONACRA, it does not take part in its metabolism in HLM. Thus, MONACRA can be utilized as a specific fluorogenic substrate of CYP4A11. We developed a robust and sensitive automated fluorimetric assay of MONACRA demethylation and used it to compare the substrate saturation profiles in seven pooled HLM preparations with the known composition of the P450 pool. These studies demonstrated a close correlation between the rate of the main kinetic phase of MONACRA metabolism and the fractional content of CYP4A11 in the P450 pool.

Indexed as

Cytochrome P-450 Enzyme SystemFluorescent DyesNaphthalenesCytochrome P-450 CYP1A2Cytochrome P-450 CYP4AFluorometryHumansKineticsMicrosomes, LiverRecombinant ProteinsSubstrate SpecificityCYP1A2 protein, humanCYP4A11 protein, humanCytochrome P-450 CYP1A2Cytochrome P-450 CYP4ACytochrome P-450 Enzyme SystemFluorescent DyesNaphthalenesRecombinant Proteins3-(6-methoxynaphthalen-2-yl)acrylic acidCYP1A2CYP4A11fluorogenic substrateshigh-throughput activity assayhuman liver microsomes

Identifiers

PMID40454940
PMCPMC12191924

What Socratic holds

Textmetadata
LicenceCC BY
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.