Biochemical characterization of a variant human medium-chain acyl-CoA dehydrogenase with a disease-associated mutation localized in the active site

Küchler, Burkhard; Abdel Ghany, Abdel Ghany; Bross, Peter; Nandy, Andreas; Rasched, Ihab; Ghisla, Sandro

Biochemical characterization of a variant human medium-chain acyl-CoA dehydrogenase with a disease-associated mutation localized in the active site

Dateien

Biochemical_characterization_of_a_variant_human_medium_chain_acyl_CoA_dehydrogenase_with_a_disease_associated_mutation_localized_in_the_active_site.pdfGröße: 183.93 KBDownloads: 431

Datum

1999

Autor:innen

Küchler, Burkhard

Abdel Ghany, Abdel Ghany

Bross, Peter

Nandy, Andreas

Rasched, Ihab

Ghisla, Sandro

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Biologie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Biochemical Journal. 1999, 337(2), pp. 225-230. ISSN 0264-6021. eISSN 1470-8728. Available under: doi: 10.1042/bj3370225

Zusammenfassung

Medium-chain acyl-CoA dehydrogenase (MCADH) deficiency, an autosomal recessive inherited disorder, is the most common genetic disorder in mitochondrial β-oxidation in humans. In addition to one prevalent disease-causing mutation (K304E), a series of rarer mutations has been reported, but none of these has yet been characterized in detail. We report here on the biochemical characterization of the purified recombinant mutant protein in which threonine is replaced by alanine at position 168 of the mature protein (T168A-MCADH). It is the first mutation to be found in patients that is located in the active site of the enzyme. Thr-168 is hydrogen-bonded to the flavin N(5) of the cofactor FAD. The thermostability of T168A-MCADH is markedly decreased compared with human wild-type MCADH (hwt-MCADH). Catalytic activity with ferricenium as acceptor is lowered by 80% and with the natural acceptor electron-transferring flavoprotein by over 90% compared with hwt-MCADH. In the mutant the extent of flavin semiquinone formed on reduction is approx. 50% that of hwt-MCADH. The pK reflected by the pH-dependence of Vmax is shifted from approx. 8.2 (hwt-MCADH) to approx. 7 (T168A-MCADH) and the rates of enzyme flavin reduction (stopped-flow measurements) are only approx. 1/10 those of the parent enzyme. These properties are discussed in the light of the possible mechanisms leading to disease in humans.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

KÜCHLER, Burkhard, Abdel Ghany ABDEL GHANY, Peter BROSS, Andreas NANDY, Ihab RASCHED, Sandro GHISLA, 1999. Biochemical characterization of a variant human medium-chain acyl-CoA dehydrogenase with a disease-associated mutation localized in the active site. In: Biochemical Journal. 1999, 337(2), pp. 225-230. ISSN 0264-6021. eISSN 1470-8728. Available under: doi: 10.1042/bj3370225

BibTex

@article{Kuchler1999Bioch-8444,
  year={1999},
  doi={10.1042/bj3370225},
  title={Biochemical characterization of a variant human medium-chain acyl-CoA dehydrogenase with a disease-associated mutation localized in the active site},
  number={2},
  volume={337},
  issn={0264-6021},
  journal={Biochemical Journal},
  pages={225--230},
  author={Küchler, Burkhard and Abdel Ghany, Abdel Ghany and Bross, Peter and Nandy, Andreas and Rasched, Ihab and Ghisla, Sandro}
}

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Universitätsbibliographie

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