Peroxidasin protein expression and enzymatic activity in metastatic melanoma cell lines are associated with invasive potential

Martina Paumann-Page (Lead / Corresponding author), Nikolaus F. Kienzl, Jyoti Motwani, Boushra Bathish, Louise Paton, Nicholas J. Magon, Benjamin Sevcnikar, Paul G. Furtmüller, Michael W. Traxlmayr, Christian Obinger, Mike R. Eccles, Christine Winterbourn

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Peroxidasin, a heme peroxidase, has been shown to play a role in cancer progression. mRNA expression has been reported to be upregulated in metastatic melanoma cell lines and connected to the invasive phenotype, but little is known about how peroxidasin acts in cancer cells. We have analyzed peroxidasin protein expression and activity in eight metastatic melanoma cell lines using an ELISA developed with an in-house peroxidasin binding protein. RNAseq data analysis confirmed high peroxidasin mRNA expression in the five cell lines classified as invasive and low expression in the three non-invasive cell lines. Protein levels of peroxidasin were higher in the cell lines with an invasive phenotype. Active peroxidasin was secreted to the cell culture medium, where it accumulated over time, and peroxidasin protein levels in the medium were also much higher in invasive than non-invasive cell lines. The only well-established physiological role of peroxidasin is in the formation of a sulfilimine bond, which cross-links collagen IV in basement membranes via catalyzed oxidation of bromide to hypobromous acid. We found that peroxidasin secreted from melanoma cells formed sulfilimine bonds in uncross-linked collagen IV, confirming peroxidasin activity and hypobromous acid formation. Moreover, 3-bromotyrosine, a stable product of hypobromous acid reacting with tyrosine residues, was detected in invasive melanoma cells, substantiating that their expression of peroxidasin generates hypobromous acid, and showing that it does not exclusively react with collagen IV, but also with other biomolecules.

Original languageEnglish
Article number102090
Number of pages12
JournalRedox Biology
Early online date4 Aug 2021
Publication statusPublished - 24 Oct 2021


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