Residue 134 determines the dimer–tetramer assembly of nucleoside diphosphate kinase from moderately halophilic bacteria

Tokunaga, Hiroko; Ishibashi, Matsujiro; Arisaka, Fumio; Arai, Shigeki; Kuroki, Ryota; Arakawa, Tsutomu; Tokunaga, Masao

doi:10.1016/j.febslet.2008.02.054

« Previous Next »FEBS Letters
Volume 582, Issue 7 , Pages 1049-1054, 2 April 2008

Residue 134 determines the dimer–tetramer assembly of nucleoside diphosphate kinase from moderately halophilic bacteria

Edited by Peter Brzezinski

Hiroko Tokunaga
- Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan
Matsujiro Ishibashi
- Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan
Fumio Arisaka
- Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B-39 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan
Shigeki Arai
- Quantum Beam Science Directorate, Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195, Japan
Ryota Kuroki
- Quantum Beam Science Directorate, Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195, Japan
Tsutomu Arakawa
- Alliance Protein Laboratories, Thousand Oaks, CA 91360, USA
Masao Tokunaga
- Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan
- Corresponding author. Fax: +81 99 285 8634.

Received 31 January 2008; received in revised form 22 February 2008; accepted 23 February 2008. published online 04 March 2008.

Abstract

Halomonas nucleoside diphosphate kinase (HaNDK) forms a dimeric assembly and Pseudomonas NDK (PaNDK) forms a tetrameric assembly. The mutation of Glu134 to Ala in HaNDK resulted in the conversion of the native dimeric structure to the tetramer assembly. Conversely, the mutation of Ala134 to Glu in PaNDK lead to the conversion from the tetramer to the dimer assembly, indicating that a single amino acid substitution at position 134 results in an alteration of the oligomeric structure of NDK. By modeling the structure of HaNDK and PaNDK based on the crystal structure of Myxococcus NDK, we showed that Glu134 exerts sufficient repulsive forces to disrupt the dimer–dimer interaction and prevent the formation of the tetramer.

Keywords: Nucleoside diphosphate kinase, Halophilic, Subunit assembly, Chimeric protein, Cross-linking