NADPH-Independent Fluorescent Probe for Live-Cell Imaging of Heme Oxygenase-1

Overview

Journal ACS Sens

Publisher American Chemical Society

Specialty Biotechnology

Date 2025 Jan 2

PMID 39745434

Authors

Liang Li

Xuanyi Lu

Qiyuan He

Chao Shu

Edward R H Walter

Lin Wang

Nicholas J Long

Lijun Jiang

Affiliations

Soon will be listed here.

Abstract

Heme oxygenase-1 (HO-1) catalyzes heme degradation on the consumption of NADPH and molecular oxygen. As an inducible enzyme, HO-1 is highly induced in various disease states, including cancer. Currently, two fluorescent probes for HO-1 have been designed based on the catalytic activity of HO-1, in which the probes serve as a substrate, so NADPH is required to enable the detection. Probes functioning in a NADPH-dependent way may influence other NADPH-consuming pathways, as all these pathways share a common NADPH pool. Here, we report the peptide-based fluorescent probe NBD-P as a simple alternative approach for HO-1 sensing. The designed probe NBD-P functions independently of the catalytic activity of HO-1, therefore enabling fast and sensitive detection of HO-1 with no requirements of other substances, including NADPH and biliverdin reductase. Moreover, it overcomes the need for a large substrate amount and long incubation time during the detection. NBD-P can be quickly taken up by cells, demonstrates an excellent colocalization with the endoplasmic reticulum (where HO-1 is mainly located), and is shown to be reliable in reporting changes in HO-1 levels in live cells. This work provides a simple alternative approach for designing HO-1 fluorescent probes, and we expect it will act as a practical tool for further studying HO-1 biology.

References

Jiang L, Lung H, Huang T, Lan R, Zha S, Chan L . Reactivation of Epstein-Barr virus by a dual-responsive fluorescent EBNA1-targeting agent with Zn-chelating function. Proc Natl Acad Sci U S A. 2019; 116(52):26614-26624. PMC: 6936348. DOI: 10.1073/pnas.1915372116. View

Funes S, Rios M, Fernandez-Fierro A, Covian C, Bueno S, Riedel C . Naturally Derived Heme-Oxygenase 1 Inducers and Their Therapeutic Application to Immune-Mediated Diseases. Front Immunol. 2020; 11:1467. PMC: 7396584. DOI: 10.3389/fimmu.2020.01467. View

Kweon M, Adhami V, Lee J, Mukhtar H . Constitutive overexpression of Nrf2-dependent heme oxygenase-1 in A549 cells contributes to resistance to apoptosis induced by epigallocatechin 3-gallate. J Biol Chem. 2006; 281(44):33761-72. DOI: 10.1074/jbc.M604748200. View

Jung N, Kim H, Kim B, Cha S, Kim G, Ha H . Evidence for heme oxygenase-1 association with caveolin-1 and -2 in mouse mesangial cells. IUBMB Life. 2003; 55(9):525-32. DOI: 10.1080/15216540310001620968. View

Wu J, Li S, Li C, Cui L, Ma J, Hui Y . The non-canonical effects of heme oxygenase-1, a classical fighter against oxidative stress. Redox Biol. 2021; 47:102170. PMC: 8577501. DOI: 10.1016/j.redox.2021.102170. View

Tagami Y, Hara Y, Murohashi K, Nagasawa R, Fujii H, Izawa A . Serum heme oxygenase-1 as a prognostic biomarker in patients with acute exacerbation of interstitial lung disease. Sci Rep. 2023; 13(1):22639. PMC: 10730846. DOI: 10.1038/s41598-023-49342-4. View

Karimi Z, Ghaffari M, Ezzati Nazhad Dolatabadi J, Dehghan P . The protective effect of thymoquinone on -butylhydroquinone induced cytotoxicity in human umbilical vein endothelial cells. Toxicol Res (Camb). 2020; 8(6):1050-1056. PMC: 7488589. DOI: 10.1039/c9tx00235a. View

Ryter S, Kvam E, Richman L, Hartmann F, Tyrrell R . A chromatographic assay for heme oxygenase activity in cultured human cells: application to artificial heme oxygenase overexpression. Free Radic Biol Med. 1998; 24(6):959-71. DOI: 10.1016/s0891-5849(97)00380-8. View

Nitti M, Ivaldo C, Traverso N, Furfaro A . Clinical Significance of Heme Oxygenase 1 in Tumor Progression. Antioxidants (Basel). 2021; 10(5). PMC: 8155918. DOI: 10.3390/antiox10050789. View

10.

Lee S, Xie J, Chen X . Peptide-based probes for targeted molecular imaging. Biochemistry. 2010; 49(7):1364-76. PMC: 2829605. DOI: 10.1021/bi901135x. View

11.

Pittala V, Salerno L, Romeo G, Modica M, Siracusa M . A focus on heme oxygenase-1 (HO-1) inhibitors. Curr Med Chem. 2013; 20(30):3711-32. DOI: 10.2174/0929867311320300003. View

12.

Jiang C, Huang H, Kang X, Yang L, Xi Z, Sun H . NBD-based synthetic probes for sensing small molecules and proteins: design, sensing mechanisms and biological applications. Chem Soc Rev. 2021; 50(13):7436-7495. PMC: 8763210. DOI: 10.1039/d0cs01096k. View

13.

Higashimoto Y, Sakamoto H, Hayashi S, Sugishima M, Fukuyama K, Palmer G . Involvement of NADPH in the interaction between heme oxygenase-1 and cytochrome P450 reductase. J Biol Chem. 2004; 280(1):729-37. DOI: 10.1074/jbc.M406203200. View

14.

Sierra E, Nutter L . A microassay for heme oxygenase activity using thin-layer chromatography. Anal Biochem. 1992; 200(1):27-30. DOI: 10.1016/0003-2697(92)90271-8. View

15.

Campbell N, Fitzgerald H, Dunne A . Regulation of inflammation by the antioxidant haem oxygenase 1. Nat Rev Immunol. 2021; 21(7):411-425. DOI: 10.1038/s41577-020-00491-x. View

16.

Taira J, Sugishima M, Kida Y, Oda E, Noguchi M, Higashimoto Y . Caveolin-1 is a competitive inhibitor of heme oxygenase-1 (HO-1) with heme: identification of a minimum sequence in caveolin-1 for binding to HO-1. Biochemistry. 2011; 50(32):6824-31. DOI: 10.1021/bi200601t. View

17.

Swiecicki J, Pisa M, Burlina F, Lecorche P, Mansuy C, Chassaing G . Accumulation of cell-penetrating peptides in large unilamellar vesicles: A straightforward screening assay for investigating the internalization mechanism. Biopolymers. 2015; 104(5):533-43. DOI: 10.1002/bip.22652. View

18.

TENHUNEN R, MARVER H, Schmid R . The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci U S A. 1968; 61(2):748-55. PMC: 225223. DOI: 10.1073/pnas.61.2.748. View

19.

Zhao Z, Xu Y, Lu J, Xue J, Liu P . High expression of HO-1 predicts poor prognosis of ovarian cancer patients and promotes proliferation and aggressiveness of ovarian cancer cells. Clin Transl Oncol. 2017; 20(4):491-499. DOI: 10.1007/s12094-017-1738-7. View

20.

Sun D, Dong G, Wu Y, Dong G, Du L, Li M . Fluorescent and theranostic probes for imaging nicotinamide phosphoribosyl transferase (NAMPT). Eur J Med Chem. 2023; 248:115080. DOI: 10.1016/j.ejmech.2022.115080. View