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Glucuronidation of d-Luciferin In Vitro: Isoform Selectivity and Kinetics Characterization

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European Journal of Drug Metabolism and Pharmacokinetics Aims and scope Submit manuscript

Abstract

Background

d-luciferin is one of the most commonly used substrates in bioluminescence imaging for real-time monitoring of sophisticated biological processes in models of human biology or disease in vitro and in vivo. d-luciferin is rapidly cleared from blood circulation after being exogenously delivered in vivo and the presence of phenolic groups indicates that glucuronide conjugation is a possible metabolic pathway for the compound.

Objectives

This study aimed to characterize the glucuronidation pathway of d-luciferin in human liver microsomes (HLM) and human intestine microsomes (HIM).

Methods

HLM and HIM were employed to catalyze the glucuronidation of d-luciferin in vitro. The activity of recombinant uridine-diphospho-glucuronosyltransferase (UGT) isoforms towards d-luciferin glucuronidation was screened. Chemical inhibition assay and kinetic analysis was combined to determine the UGT isoforms responsible for the formation of d-luciferin glucuronide in HLM and HIM.

Results

d-luciferin could be catalyzed to form one mono-glucuronide which was characterized as 6′-O-glucuronide in HLM and HIM. UGT1A1, 1A3, 1A6, 1A8, 1A9 and 1A10 participated in the formation of d-luciferin glucuronide, with UGT1A1 exhibiting the highest catalytic activity. Both chemical inhibition assays and kinetic analysis showed that UGT1A1 and UGT1A3 played important roles in d-luciferin-6′-O-glucuronidation in HLM and HIM, with UGT1A6 also giving a non-negligible contribution to this biotransformation in HLM.

Conclusions

UGT1A1, UGT1A3 and UGT1A6 were responsible for 6′-O-glucuronidation of d-luciferin in HLM, while UGT1A1 and UGT1A3 were the major contributors to this biotransformation in HIM.

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Acknowledgements

We thank Dr. Guangbo Ge and Dr. Yong Liu for their kind help in writing and editing the manuscript.

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Authors and Affiliations

  1. School of Life Science and Medicine, Dalian University of Technology, Panjin, China

    Yangliu Xia & Huilin Pang

Authors
  1. Yangliu Xia
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  2. Huilin Pang
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Corresponding author

Correspondence to Yangliu Xia.

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Funding

This work was supported by the National Natural Science Foundation of China (81603187), and the Fundamental Research Funds for the Central Universities (DUT18LK39).

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Ethics Approval

This study was performed in accordance with the Guidelines for Care and Use of Laboratory Animals of Shanghai University of Traditional Chinese Medicine and the experiments were approved by the Institutional Animal Ethics Committee, Science and Technology Commission of Shanghai Municipality with the reference number SYXK2014-0008.

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Xia, Y., Pang, H. Glucuronidation of d-Luciferin In Vitro: Isoform Selectivity and Kinetics Characterization. Eur J Drug Metab Pharmacokinet 44, 549–556 (2019). https://doi.org/10.1007/s13318-019-00549-9

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  • DOI: https://doi.org/10.1007/s13318-019-00549-9

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