Publikation:

Location of the Active Site and Proposed Catalytic Mechanism of Pterin-4A-Carbinolamine Dehydratase

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1996

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Köster, Sandra
Stier, Gunter
Ficner, Ralf
Hölzer, Manuela
Curtius, Hans-Christoph
Suck, Dietrich

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Published

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European Journal of Biochemistry. 1996, 241(3), pp. 858-864. ISSN 0014-2956. eISSN 1432-1033. Available under: doi: 10.1111/j.1432-1033.1996.00858.x

Zusammenfassung

Based on the recently solved three-dimensional structure of pterin-4a-carbinolamine dehydratase from rat/human liver the involvement of the proposed active-site residues Glu57, Asp60, His61, His62, Tyr69, His79, Arg87 and Asp88 was examined by site-directed mutagenesis. Most of the mutants showed reduced activity, and only the Glu57→Ala mutant and the His61→Ala, His62→Ala double mutant were fully devoid of activity. The dissociation constants of quinonoid 6,6-dimethyl-7,8-dihydropterin were significantly increased for binding to the Glu57→Ala, His61→Ala, His62→Ala single mutants and the His61→Ala, His62→Ala double mutant, confirming that His61 and His62 are essential for substrate binding and catalysis. The mechanism of dehydration is proposed to involve base catalysis at the N(5)-H group of the substrate by His61.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie

Schlagwörter

hepatocyte nuclear factor-1α-dimerization cofactor, dehydratase, tetrahydrobiopterin, phenylalanine hydroxylase

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ISO 690KÖSTER, Sandra, Gunter STIER, Ralf FICNER, Manuela HÖLZER, Hans-Christoph CURTIUS, Dietrich SUCK, Sandro GHISLA, 1996. Location of the Active Site and Proposed Catalytic Mechanism of Pterin-4A-Carbinolamine Dehydratase. In: European Journal of Biochemistry. 1996, 241(3), pp. 858-864. ISSN 0014-2956. eISSN 1432-1033. Available under: doi: 10.1111/j.1432-1033.1996.00858.x
BibTex
@article{Koster1996Locat-8396,
  year={1996},
  doi={10.1111/j.1432-1033.1996.00858.x},
  title={Location of the Active Site and Proposed Catalytic Mechanism of Pterin-4A-Carbinolamine Dehydratase},
  number={3},
  volume={241},
  issn={0014-2956},
  journal={European Journal of Biochemistry},
  pages={858--864},
  author={Köster, Sandra and Stier, Gunter and Ficner, Ralf and Hölzer, Manuela and Curtius, Hans-Christoph and Suck, Dietrich and Ghisla, Sandro}
}
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