Description
General description
Research area: Cell signaling. Phosphodiesterase (PDE) II belongs to the PDE superfamily and includes subtypes from one to 11. They exist as a dimer and comprise of N-terminal regulatory domain, C-terminal prenylated domain, and a Zn2+containing catalytic domain. The bovine spleen PDE2 corresponds to a molecular weight of 65 kDa. It requires divalent cations for its enzymatic activity and has an optimum pH of 7.5.
Application
Phosphodiesterase II from bovine spleen has been used in the:
excision of pyridyloxobutyl (POB) base adducts from DNA
digestion of DNA prior to cycloadenosine enrichment
hydrolysis of DNA from blue mussels gill tissue into deoxyribonucleoside 3′-monophosphates
Phosphodiesterase (PDE) is any enzyme that is used to break phosphodiester bonds. It is a membrane-bound glycoprotein that is used to catalyze the hydrolysis of various nucleotide polyphosphates. Phosphodiesterase II has been used in the enzymatic digestions of purified proteins such as the P8-dGMP complex. Bovine spleen phosphodiesterase has been used to digest N-cadherin. Phosphodiesterase II from bovine is suitable for sequential and end-group analysis of poly and oligonucleotides. It may be used in kinetic studieson substrate degradation of macro-molecules.
The product has been used in the characterization of polynucleotide chain length, base composition, and identity of terminal nucleotide. The enzyme has also been used in excision of pyridyloxobutyl (POB) base adducts from DNA. Furthermore, it has been used along with micrococcal endonuclease to hydrolyze purified DNA to 3 -nucleoside monophosphates.
Biochem/physiol Actions
Phosphodiesterase (PDE) breaks phosphodiester bonds. 3-Isobutyl-methylxanthine (IBMX) is a potential PDE2 inhibitor.
Hydrolyzes RNA, RNA-Core, 3′-alkyl- and 3′-aryl-nucleoside phosphates, and polydeoxyribonucleotides with 3′-phosphate end groups to 3′-mononucleotides.
Polynucleotides having 5′-phosphomonoester end groups are not attacked.
The enzyme acts on poly(A), poly(U), and poly(I). Native DNA and poly(C) are quite resistant to the action of this enzyme.
Analysis Note
Protein determined by biuret
Other Notes
One unit will produce acid soluble nucleotides equivalent to a ΔA260 of 16 in 30 min at pH 6.5 at 37 °C, in a 2.0 mL reaction mixture. Substrate: RNA-Core. Actual A260 is measured on the supernatant after precipitation of the unhydrolyzed RNA with uranyl acetate-perchloric acid reagent.
