A rapid and sensitive colorimetric assay for the determination of adenosine kinase activity

Biochem Biophys Res Commun. 2018 Jul 12;502(2):250-254. doi: 10.1016/j.bbrc.2018.05.152. Epub 2018 May 26.

Abstract

Adenosine kinase (ADK) plays an important role in the growth and development of organisms. A convenient, quick, reliable, sensitive and low-cost assay for ADK activity is of great significance. Here, we found the reaction system with bromothymol blue as the pH indicator had a maximum absorption peak at 614 nm. The absorbance change in 614 nm was positively correlated with the generated hydrogen ions in the reaction catalyzed by ADK. Then, we demonstrated this assay was feasible for ADK activity. Further, we analyzed the effects of buffer, bromothymol blue concentrations on the sensitivity of the assay, and investigated the sensitivity of ADK contents and adenosine concentration on the assay. Finally, we calculated the Km and Vmax of ADK from Bombyx mori with this assay. Our results suggested this assay was quick, convenient, reliable, sensitive and economic for the activity of ADK. It is an excellent alternative for the conventional ADK assays.

Keywords: Adenosine kinase; Bromothymol blue; Convenient; Enzymatic activity assay; Quick; Sensitive.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Kinase / analysis*
  • Adenosine Kinase / chemistry
  • Adenosine Kinase / metabolism
  • Animals
  • Bombyx / enzymology
  • Bromphenol Blue
  • Bromthymol Blue
  • Buffers
  • Colorimetry / methods*
  • Colorimetry / statistics & numerical data
  • Coloring Agents
  • Feasibility Studies
  • Hydrogen-Ion Concentration
  • Kinetics
  • Spectrophotometry

Substances

  • Buffers
  • Coloring Agents
  • Bromphenol Blue
  • Adenosine Kinase
  • Bromthymol Blue