Characterization of the Asp94 and Glu242 mutants in myeloperoxidase, the residues linking the heme group via ester bonds

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Kooter, Ingeborg M. and Moguilevsky, Nicole and Bollen, Alex and Sijtsema, Nanna M. and Otto, Cees and Dekker, Henk L. and Wever, Ron (1999) Characterization of the Asp94 and Glu242 mutants in myeloperoxidase, the residues linking the heme group via ester bonds. European Journal of Biochemistry, 264 (1). pp. 211-217. ISSN 0014-2956

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Abstract:The heme group of all mammalian peroxidases is covalently linked to the protein matrix via two esterbonds, as we have recently shown by Fourier transform infrared (FTIR) difference spectroscopy [Kooter, I.M., Pierik, A.J., Merkx, M., Averill, B.A., Moguilevsky, N., Bollen, A. & Wever, R. (1997) J. Am. Chem. Soc.119, 11542–11543]. We have examined the effects of mutation of Asp94 and Glu242, responsible for those ester bonds in myeloperoxidase, on the spectroscopic properties and catalytic activity of this enzyme. Mutation of Asp94 in myeloperoxidase results in two species. The first species has spectroscopic characteristics similar to that of wild-type myeloperoxidase. The second species has spectroscopic characteristics similar to that of Met243→Gln mutant, and it is therefore concluded that, besides loss of the ester bond involving Asp94, this species also has lost the sulfonium ion linkage that is also characteristic of myeloperoxidase. The Asp94→Asn mutant still has about 30% residual peroxidase activity while for the Asp94→Val mutant only a few percentage activity is left. When Glu242 is mutated the sulfonium ion linkage is not affected, but this residue together with its neighbouring residue Met243, according to resonance Raman spectra, is responsible for the low symmetry of the heme group. Mutation of either of these residues results in loss of the bowed distortion from the planar conformation, and in a heme group with higher symmetry. For the Glu242→Gln mutant 8% residual peroxidase activity is found.
Item Type:Article
Copyright:© 1999 Wiley InterScience
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Science and Technology (TNW)
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Link to this item:http://purl.utwente.nl/publications/71491
Official URL:http://dx.doi.org/10.1046/j.1432-1327.1999.00606.x
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Metis ID: 128421