Non disponible en dehors du Royaume-Uni et de l'Irlande

Analysis Note

Protein determined by biuret

Application

Phosphodiesterase (PDE) is any enzyme that is used to breaks 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.

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.

Phosphodiesterase II from bovine spleen has been used in the: excision of pyridyloxobutyl (POB) base adducts from DNAdigestion of DNA prior to cycloadenosine enrichment hydrolysis of DNA from blue mussels gill tissue into deoxyribonucleoside 3′-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.

General description

Phosphodiesterase (PDE) II belong to the PDE superfamily and includes subtypes from one to 11. They exists as a dimer and comprise a 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.

Packaging

10 units in glass bottle

25 units in poly bottle

Unit Definition

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.