Structure-function studies of human deoxyhypusine synthase: identification of amino acid residues critical for the binding of spermidine and NAD

The Biochemical Journal
C H LeeM H Park

Abstract

Deoxyhypusine synthase catalyses the first step in the biosynthesis of hypusine [N(epsilon)-(4-amino-2-hydroxybutyl)lysine]. The crystal structure of human deoxyhypusine synthase in complex with NAD revealed four NAD-binding sites per enzyme tetramer, and led to a prediction of the spermidine-binding pocket. We have replaced each of the seven amino acid residues at the predicted spermidine-binding site, and eleven residues that contact NAD, on an individual basis with alanine. Of the amino acid residues at the spermidine site, substitution of Asp-243, Trp-327, His-288, Asp-316 or Glu-323 with alanine caused an almost complete loss of spermidine binding and enzyme activity; only the mutation Tyr-305-->Ala showed partial binding and activity. His-288-->Ala was also deficient in terms of binding NAD. NAD binding was significantly reduced in all of the NAD-site mutant enzymes, except for Glu-137-->Ala, which showed a normal binding of NAD, but was totally lacking in spermidine binding. Of the NAD-site mutant enzymes, Asp-342-->Ala, Asp-313-->Ala and Asp-238-->Ala displayed the lowest binding of NAD. These enzymes and His-288Ala also showed a reduced binding of spermidine, presumably because spermidine binding is dependent on NAD. T...Continue Reading

Citations

Sep 28, 2015·Biochimie·Laura Itzel Quintas-GranadosMaría Elizbeth Álvarez-Sánchez
Sep 10, 2004·DNA Sequence : the Journal of DNA Sequencing and Mapping·Jenq-Kuen HuangLisa Wen
Apr 3, 2009·Electrophoresis·Xing-Zheng WuYoshie Yamaguchi
Sep 28, 2018·The Journal of Biological Chemistry·Myung Hee Park, Edith C Wolff
Nov 3, 2009·The Journal of Biological Chemistry·Bhavna ChawlaRentala Madhubala
Jun 4, 2015·The Journal of Biological Chemistry·Nora PällmannStefan Balabanov
Mar 7, 2020·Journal of Medicinal Chemistry·Yuta TanakaShinichi Imamura
Mar 25, 2004·Protein Expression and Purification·Jenq-Kuen HuangLisa Wen

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