Exploring the Chemical Space of Proluciferins as Probe Substrates for Human Cytochrome P450 Enzymes

Zhao, Jie; Zhang, Xue; Wang, Yueyin; Huang, Huimin; Sharma, Shraddha; Sharma, Sangeeta Shrestha; Wolf, Clemens Alexander; Liu, Sijie; Wolber, Gerhard; Sorensen, Erik J.; Bureik, Matthias

doi:10.1007/s12010-022-04184-0

Cited by 3 publications

(5 citation statements)

References 39 publications

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“…The presence of a chlorine group in luciferin CEE and a fluorine one in luciferin FEE might cause various interactions that involve the active site of the human CYP2C9, which could explain the activities that we have measured. It should also be noted that the activity of the human CYP2C9 against luciferin CEE in S. cerevisiae was close to that reported by Promega [ 35 ], whereas no activity was found in S. pombe [ 27 , 35 , 36 ]. Finally, we observed also a rather high activity against Luc-H, although much lower than that previously reported in S. pombe cells [ 36 , 37 ] (see Fig.…”

Section: Resultssupporting

confidence: 81%

“…We tested the activity of the human CYP2C9 on ten different luciferin substrates (see Table S4 ). Except for Luc-H (from Promega), these substrates were previously synthesized in-house (and used in S. pombe enzyme bags containing various human P450s) [ 17 , 26 , 27 ]. S. cerevisiae strain byMM1961—and its control byMM1959 (expressing only human CPR)—were employed to analyze the biotransformation of the above-mentioned luciferin substrates.…”

Section: Resultsmentioning

confidence: 99%

“…CORM-401 stock solution was heated at 37 °C just before adding it to a reaction mixture with substrates and it was always protected from light. Luc-BE, Luc-2FBE, Luc-3FBE, Luc-CEE, Luc-FEE, Luc-EE, Luc-FuBE, Luc-PE, and Luc-CPE were synthesized by our collaborative group [ 17 , [26] , [27] , [28] ]. All other chemicals and reagents used in this work are of the highest available grade.…”

Section: Methodsmentioning

confidence: 99%

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Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells

Ashraf,

Bureik,

Marchisio

2024

Synthetic and Systems Biotechnology

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Section: Resultssupporting

confidence: 81%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells

Ashraf,

Bureik,

Marchisio

2024

Synthetic and Systems Biotechnology

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“…Therefore, it was the aim of the present study to determine the effect of co-expression of each one of the 19 human members of the UGT families 1 and 2 on the activity of three human CYPs. In this way, two of the most important drug metabolizing CYPs (CYP2C9 and CYP2D6) could be tested with CYP4Z1 as an internal control (as it is not known to have a function in xenobiotic biotransformation, and many of the marketed and in-house synthesized luciferin probe substrates are metabolized by this enzyme [23][24][25][26], which makes it a reliable control for bioluminescent assay in CYP metabolism).…”

Section: Introductionmentioning

confidence: 99%

Mutual Influence of Human Cytochrome P450 Enzymes and UDP-Glucuronosyltransferases on Their Respective Activities in Recombinant Fission Yeast

Sharma

Zhao

et al. 2023

Biomedicines

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Cytochromes P450 (CYPs) and UDP-glucuronosyltransferases (UGTs) are the most important human drug metabolizing enzymes, but their mutual interactions are poorly understood. In this study, we recombinantly co-expressed of each one of the 19 human members of the UGT families 1 and 2 with either CYP2C9, CYP2D6, or CYP4Z1 in fission yeast. Using these strains, we monitored a total of 72 interactions: 57 cases where we tested the influence of UGT co-expression on CYP activity and 15 cases of the opposite approach. In the majority of cases (88%), UGT co-expression had a statistically significant (p < 0.05) effect on P450 activity (58% positive and 30% negative). Strong changes were observed in nine cases, including one case with an activity increase by a factor of 23 (CYP2C9 activity in the presence of UGT2A3) but also four cases with a complete loss of activity. When monitoring the effect of CYP co-expression on the activity of five UGTs, activity changes were generally not so pronounced and, if observed, always detrimental. UGT2B7 activity was not influenced by CYP co-expression, while the other UGTs were affected to varying degrees. These data suggest the notion that mutual influence of CYPs and UGTs on each other’s activity is a widespread phenomenon.

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“…Some of these compounds (preproluciferins) need an additional reaction step for conversion into luciferin proper. In recent years we have significantly expanded the set of proluciferin probe substrates in several studies [11][12][13]. Until the present study, Luciferin-H and Luciferin-ME were the only known CYP2A7 substrates, and testing additional proluciferin probes was an obvious choice to start the current investigation.…”

Section: Introductionmentioning

confidence: 98%

Identification of New Substrates and Inhibitors of Human CYP2A7

Ashraf,

Liu,

Wolf

et al. 2024

Molecules

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CYP2A7 is one of the most understudied human cytochrome P450 enzymes and its contributions to either drug metabolism or endogenous biosynthesis pathways are not understood, as its only known enzymatic activities are the conversions of two proluciferin probe substrates. In addition, the CYP2A7 gene contains four single-nucleotide polymorphisms (SNPs) that cause missense mutations and have minor allele frequencies (MAFs) above 0.5. This means that the resulting amino acid changes occur in the majority of humans. In a previous study, we employed the reference standard sequence (called CYP2A7*1 in P450 nomenclature). For the present study, we created another CYP2A7 sequence that contains all four amino acid changes (Cys311, Glu169, Gly479, and Arg274) and labeled it CYP2A7-WT. Thus, it was the aim of this study to identify new substrates and inhibitors of CYP2A7 and to compare the properties of CYP2A7-WT with CYP2A7*1. We found several new proluciferin probe substrates for both enzyme variants (we also performed in silico studies to understand the activity difference between CYP2A7-WT and CYP2A7*1 on specific substrates), and we show that while they do not act on the standard CYP2A6 substrates nicotine, coumarin, or 7-ethoxycoumarin, both can hydroxylate diclofenac (as can CYP2A6). Moreover, we found ketoconazole, 1-benzylimidazole, and letrozole to be CYP2A7 inhibitors.

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Exploring the Chemical Space of Proluciferins as Probe Substrates for Human Cytochrome P450 Enzymes

Cited by 3 publications

References 39 publications

Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells

Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells

Mutual Influence of Human Cytochrome P450 Enzymes and UDP-Glucuronosyltransferases on Their Respective Activities in Recombinant Fission Yeast

Identification of New Substrates and Inhibitors of Human CYP2A7

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