Biochemistry (Moscow)

, Volume 73, Issue 1, pp 38–43

Identification of a novel nuclear-localized adenylate kinase from Drosophila melanogaster

Article

Abstract

As a step to further understand the role of adenylate kinase (AK) in the energy metabolism network, we identified, purified, and characterized a previously underscribed adenylate kinase in Drosophila melanogaster. The cDNA encodes a 175-amino acid protein, which shows 47.85% identity in 163 amino acids to human AK6. The recombinant protein was successfully expressed in Escherichia coli BL21 (DE3) strain. Characterization of this protein by enzyme activity assay showed adenylate kinase activity. AMP and CMP were the preferred substrates, and UMP can also be phosphorylated to some extent, with ATP as the best phosphate donor. Subcellular localization study showed a predominantly nuclear localization. Therefore, based on the substrate specificity, the specific nuclear localization in the cell, and the sequence similarity with human AK6, we named this novel adenylate kinase identified from the fly DAK6.

Key words

cloning protein expression adenylate kinase activity nuclear localization 

Abbreviations

AK

adenylate kinase

DAPI

4,6-diamidino-2-phenylindole

GFP

green fluorescent protein

IPTG

isopropyl β-thiogalactopyranoside

NLS

nuclear localization sequence

NMPK

nucleoside monophosphate kinases.

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Copyright information

© Pleiades Publishing, Ltd. 2008

Authors and Affiliations

  • Geng Meng
    • 1
    • 2
  • Ruitong Zhai
    • 2
  • Bin Liu
    • 2
  • Xiaofeng Zheng
    • 1
    • 2
  1. 1.National Laboratory of Protein Engineering and Plant Genetic EngineeringBeijingChina
  2. 2.Department of Biochemistry and Molecular Biology, College of Life SciencesPeking UniversityBeijingChina

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