Crystal structures of blasticidin S deaminase (BSD): Implications for dynamic properties of catalytic zinc

Takashi Kumasaka, Masaki Yamamoto, Makio Furuichi, Masayoshi Nakasako, Aik Hong Teh, Makoto Kimura, Isamu Yamaguchi, Tatzuo Ueki

研究成果: Article

14 引用 (Scopus)

抜粋

The set of blasticidin S (BS) and blasticidin S deaminase (BSD) is a widely used selectable marker for gene transfer experiments. BSD is a member of the cytidine deaminase (CDA) family; it is a zinc-dependent enzyme with three cysteines and one water molecule as zinc ligands. The crystal structures of BSD were determined in six states (i.e. native, substrate-bound, product-bound, cacodylate-bound, substrate-bound E56Q mutant, and R90K mutant). In the structures, the zinc position and coordination structures vary. The substrate-bound structure shows a large positional and geometrical shift of zinc with a double-headed electron density of the substrate that seems to be assigned to the amino and hydroxyl groups of the substrate and product, respectively. In this intermediate-like structure, the steric hindrance of the hydroxyl group pushes the zinc into the triangular plane consisting of three cysteines with a positional shift of ∼0.6 Å, and the fifth ligand water approaches the opposite direction of the substrate with a shift of 0.4 Å. Accordingly, the zinc coordination is changed from tetrahedral to trigonal bipyramidal, and its coordination distance is extended between zinc and its intermediate. The shift of zinc and the recruited water is also observed in the structure of the inactivated E56Q mutant. This novel observation is different in two-cysteine cytidine deaminase Escherichia coli CDA and might be essential for the reaction mechanism in BSD, since it is useful for the easy release of the product by charge compensation and for the structural change of the substrate.

元の言語English
ページ(範囲)37103-37111
ページ数9
ジャーナルJournal of Biological Chemistry
282
発行部数51
DOI
出版物ステータスPublished - 2007 12 21

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Kumasaka, T., Yamamoto, M., Furuichi, M., Nakasako, M., Teh, A. H., Kimura, M., Yamaguchi, I., & Ueki, T. (2007). Crystal structures of blasticidin S deaminase (BSD): Implications for dynamic properties of catalytic zinc. Journal of Biological Chemistry, 282(51), 37103-37111. https://doi.org/10.1074/jbc.M704476200