Autocatalytic formation of a covalent link between tryptophan 41 and the heme in ascorbate peroxidase.

Journal article

Electronic spectroscopy, HPLC analyses, and mass spectrometry (MALDI-TOF and MS/MS) have been used to show that a covalent link from the heme to the distal Trp41 can occur on exposure of ascorbate peroxidase (APX) to H2O2 under noncatalytic conditions. Parallel analyses with the W41A variant and with APX reconstituted with deuteroheme clearly indicate that the covalent link does not form in the absence of either Trp41 or the heme vinyl groups. The presence of substrate also precludes formation of the link. Formation of a protein radical at Trp41 is implicated, in a reaction mechanism that is analogous to that proposed [Ghiladi, R. A., et al. (2005) Biochemistry 44, 15093-15105] for formation of a covalent Trp-Tyr-Met link in the closely related catalase peroxidase (KatG) enzymes. Collectively, the data suggest that radical formation at the distal tryptophan position is not an exclusive feature of the KatG enzymes and may be used more widely across other members of the class I heme peroxidase family.

Authors: Pipirou, Z; Bottrill, A; Metcalfe, C; Mistry, S; Badyal, S

• Publication status: Published
• Journal: Biochemistry
• Volume: 46
• Issue: 8
• Pages: 2174-2180
• Publication date: 2007-02-01
• DOI: 10.1021/bi062274q
• EISSN: 1520-4995
• ISSN: 0006-2960
• Language: English
• Keywords: Soybeans; Ascorbate Peroxidases; Bacterial Proteins; Deuteroporphyrins; Recombinant Proteins; Tandem Mass Spectrometry; Heme; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectrum Analysis; Chromatography, High Pressure Liquid; Hydrogen Peroxide; Cytochrome-c Peroxidase; Catalase; Oxidation-Reduction; Peroxidases; Tryptophan