Phosphodiesterase 5A, Recombinant, Mouse (Phosphodiesterase 5A cGMP-specific, PDE5A, CGB-PDE, cGMP-b

The cyclic monophosphate nucleotides (cyclic adenosine monophosphate [cAMP] and cyclic guanosine monophosphate [cGMP]) are found ubiquitously in mammalian cells and act as second messenger transducers to effect the intracellular actions of a variety of G protein coupled receptors (GPCRs) for hormones, cytokines, and neurotransmitters. Cyclic nucleotides are important intracellular second messengers which play important role in a variety of signal transduction process. The cyclic nucleotides are hydrolyzed and compartmentalized by a family of enzymes called phosphodieterases. One of the many phosphodiesterases that compartmentalized and hydrolyze cGMP in various tissues is phosphodiesterase type 5A (PDE5A). The cGMP is involved in nitric oxide signaling as well as cell signaling associated with natriuretic peptides and gulanylins. Some of the intracellular biding cites for the cGMP include cyclic nucleotide gated ion channels, cGMP dependent protein kinases, and cyclic GMP biding phosphodiesterases (cGB PDEs). The cGB PDEs include PDE2, PDE3, PDE5, PDE5 and PDE10, the members of these families contain various structural and functional motifs that are conserved. Most of these proteins contain dimeric subunits that contain a highly conserved cGMP binding site and a phosphodiesterase catalytic site. The cGMP specific phosphodiesterase type 5A (PDE5A) family is comprised of 2 genes (PDE5A and PDE5B) each with multiple splice variants generated by RNA splicing and use of alternate initiation sites. The PDE5 is highly expressed in aorta and lungs, intestine, kidney adrenal gland cerebellum and cerebrum. In cerebellum the PDE5 is highly expressed during neonatal development in Purkinge cells layer. The PDE5 is also abundant in vascular smooth muscle regulating cGMP levels and vascular smooth muscle tonicity. In corpus cavernosum inhibition of PDE5 by sildenafil corrects erectile dysfunctions. The nitric oxide donor sodium notropurside (SNP) stimulate PDE5 activity by cGMP dependent kinase phosphorylation. PDE5A1 has 875 amino acids (99.5 kDa). The amino terminal 142aa of the PDE5 gene showed no sequence homology with other PDEs and also contained serine 92 that is phosphorylated by cGMP kinases. There is growing evidence that supports important roles for PDE5A in both the vasculature and heart. , Full length recombinant mouse PDE5A with an N-terminal GST tag, expressed in an Baculovirus infected Sf9 cell expression system. Specific Activity: As reported, Unit Definition: 1 unit is defined as the amount of enzyme that will convert 1 pmole of 3', 5'-cGMP to 5' GMP per minute at 37°C, Assay Conditions: 10mM Tris-HCl, pH 7.4, 10mM MgCl2, 1mM MnCl2, 0.1mg/ml BSA, 200uM cGMP, 2.5kU 5' nucleotidase, 37°C, 20 minutes. Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. Other applications not tested. Recommended Dilutions: Optimal dilution determined by the researcher. Storage and Stability: Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.