PDE1 enzyme product blog
Tags: Enzyme; Phosphodiesterase 1; PDE1 enzyme; PDE1;
The PDE1 n/a (Catalog #MBS653995) is an Enzyme produced from Crotalus adamanteus venom (Eastern Diamondback rattlesnake) and is intended for research purposes only. The product is available for immediate purchase.The PDE1 n/a product has the following accession number(s) (GI #148539632) (NCBI Accession #NP_001091918.1). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful.
To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
Venom exonuclease (Phosphodiesterase I) successively hydrolyzes 5\'-mononucleotides from 3\'-hydroxy-terminated ribo-and deoxyribo-oligonucleotides. The enzyme has been widely utilized as a tool for structural and sequence studies of nucleic acids. The enzyme has been purified with endonuclease activity being eliminated as well as 5\'-nucleotidase and nonspecific monophosphatase. It is nonspecific with respect to base or sugar moieties of nucleotides. A variety of synthetic substrates are hydrolyzed. The exonuclease will not recognize nucleoside units in the syn conformation. ADP-ribosylated proteins are cleaved at the pyrophosphate linkages by venom phosphodiesterase to yield phosphoribosyl-AMP.
Activity: 95.4 u/mg dry weight, 125 u/vial
Absorbance (A280): 1.30
Nucleotidase: 1.44%
Optimum pH: 9.8-10.4
Inhibitors: Reducing agents such as glutathione, cysteine and ascorbic acid. It is completely inhibited by 5mM EDTA while ATP, ADP and AMP are partial inhibitors.
Activators: The enzyme has an absolute requirement for Mg2+ indicates an optimum concentration of 15mM.
Unit Definition: 1 unit hydrolyzes 1 umole of p-nitrophenyl thymidine-5-phosphate per minute at 25ºC, pH 8.9. Protocol: Assay:
Method:
The assay is essentially that of Razell and Khorana (1959) where the reaction velocity is determined by an increase in absorbance at 400nm resulting from the hydrolysis of p-nitrophenyl thymidine -5\'-phosphate. One unit hydrolyzes one micromole of p-nitrophenyl thymidine-5\'-phosphate per minute at pH 8.9 and 25�C under the specified conditions.
Reagents:
-0.11M Tris-HCl buffer, pH 8.9, with 0.11M sodium chloride and 15 mM MgCl2 (Tris*Salts buffer)
- 5 mM p-nitrophenyl thymidine-5\'-phosphate.
Note: The purity of commercial preparations varies somewhat and should be considered in preparing this reagent.
Enzyme:
Dissolve at 1 mg/ml in Tris*Salts buffer to obtain a rate of 0.02-0.04 ΔA/minute.
Protocol:
1. Set spectrophotometer at 400nm and 25�C.
2. Pipette into microcuvettes as follows:
Tris*Salts buffer 0.9 ml; 5 mM p-nitrophenyl thymidine-5\'-phosphate 0.1 ml.
3. Incubate cuvettes in spectrophotometer for 3-5 minutes to reach temperature equillibrium and establish blank rate, if any. Add 10ul of diluted enzyme and record increase in A400 for 3-5 minutes. The reaction remains linear until A400 reaches about 1.2. Calculate ΔA400/minute from initial linear portion of absorbance curve.
Calculation:
Units/mg = ΔA400/min/16* x mg enzyme/ml reaction mixture
*16 is the extinction coefficient of p-nitrophenol determined in the laboratory under these conditions. The following patways have been known to be associated with this gene.