PDE7A1 hydrolyzes cCMP

FEBS Lett. 2014 Sep 17;588(18):3469-74. doi: 10.1016/j.febslet.2014.08.005. Epub 2014 Aug 13.

Abstract

The degradation and biological role of the cyclic pyrimidine nucleotide cCMP is largely elusive. We investigated nucleoside 3',5'-cyclic monophosphate (cNMP) specificity of six different recombinant phosphodiesterases (PDEs) by using a highly-sensitive HPLC-MS/MS detection method. PDE7A1 was the only enzyme that hydrolyzed significant amounts of cCMP. Enzyme kinetic studies using purified GST-tagged truncated PDE7A1 revealed a cCMP KM value of 135 ± 19 μM. The Vmax for cCMP hydrolysis reached 745 ± 27 nmol/(minmg), which is about 6-fold higher than the corresponding velocity for adenosine 3',5'-cyclic monophosphate (cAMP) degradation. In summary, PDE7A is a high-speed and low-affinity PDE for cCMP.

Keywords: Cyclic CMP; Cyclic nucleotides; Enzyme kinetics; HPLC–MS; Phosphodiesterase; Second messenger.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cyclic AMP / chemistry
  • Cyclic CMP / chemistry*
  • Cyclic Nucleotide Phosphodiesterases, Type 7 / antagonists & inhibitors
  • Cyclic Nucleotide Phosphodiesterases, Type 7 / chemistry*
  • Humans
  • Hydrolysis
  • Kinetics
  • Nitro Compounds / chemistry
  • Phosphodiesterase Inhibitors / chemistry
  • Second Messenger Systems
  • Sf9 Cells
  • Spodoptera
  • Substrate Specificity
  • Sulfonamides / chemistry

Substances

  • 3-(N,N-dimethylsulfonamido)-4-methyl-nitrobenzene
  • Nitro Compounds
  • Phosphodiesterase Inhibitors
  • Sulfonamides
  • Cyclic CMP
  • Cyclic AMP
  • Cyclic Nucleotide Phosphodiesterases, Type 7
  • PDE7A protein, human