Cyclic adenosine monophosphate (cAMP) is an important mediator for the activation of signaling cascades
The C8-cAMP caproKit improves the understanding of cAMP/PKA/AKAPs-dependent signal transduction
Cyclic adenosine monophosphate (cAMP) is a ubiquitous second messenger and used to relay extracellular signals to intracellular effectors, to activate proteins or to change membrane trafficking (1). cAMP is synthesized by adenylylcyclase using ATP as substrate upon an activation of e.g. hormone receptors like glucagone or adrenaline. A major target of cAMP is protein kinase A (PKA) which is a key player for the activation of several other proteins. PKA and other cAMP-dependent proteins phosphorylate downstream targets which consequently results in an alteration of metabolism, signal transduction, differentiation, memory or apoptosis. A deregulation of cAMP or an aberrant activation of cAMP-binding transcription factors seems to be associated with tumor burden, neuronal degradation, or developmental abnormalities (2).
Despite the tremendous importance of cAMP-dependent signaling cascades, little is known about the temporal and spatial pattern of its synthesis and mode of function. To aid the understanding of these processes caprotec™ has developed C8-cAMP caproKits™ to isolate cAMP-binding proteins.
The Capture Compound™ scaffold is attached to the C8 position of cAMP, as many cAMP-binding proteins tolerate modifications at this position.
The C8-cAMP caproKit™ includes the C8-cAMP Capture Compound, C8-cAMP competitor, all buffers, positive control protein, and streptavidin magnetic beads.
For profiling the whole cAMP-interacting proteome we recommend to use the cAMP caproKit™ complete.
Please note, that CCMS reagents (caproKits™) will perform best when used with the optimized equipment (caproBox™ and caproMag™).
For the most reproducible results it is recommended to standardize parameters such as UV wavelength, distance between sample and light source, temperature and incubation time. This can best be accomplished using a caproBox™ for cross-linking.
If you are using the proprietary CCMS technology for the first time, caprotec offers a convenient CCMS Starter Kit including a free choice of 3 independent caproKits™, Biotin Capping Kit, caproBox™, and caproMag™. Please feel free to contact us for any additional questions related to using caprotec products!
A major advantage of C8-cAMP Capture Compounds is the specific isolation of the cAMP-dependent protein kinase A (PKA) and interacting proteins (A-kinase anchoring proteins, AKAPs). Most AKAPs bind to the regulatory subunit of PKA and direct the kinase to discrete cellular locations. Anchored PKA has been shown to play important roles in various cellular processes, e.g. gene transcription and ion-channel modulation (3). More than 70 different AKAPs have been described and a large number is hypothesized but unconfirmed (4). The use of C8-cAMP caproKits aids in the identification of proteins which belong to the cAMP/PKA/AKAP interactome.
caproKits allow for discovering, isolating, and profiling members of functional protein families within a variety of biological samples. Downstream of this process the proteins can be subjected to tryptic digestion, MS or gel electrophoresis.
2) B.J.B. Simpson, A.D. Ramage, M.J. Hulme, D.J. Burns, D. Katsaros, S.P. Langdon, W.R. Miller, (1996) Cyclic adenosine 3´,5´-monophosphate-binding proteins in human ovarian cancer: Correlation with clinicopahtological features; Clin. Cancer Res. (2); 201-206.
3) M. Wang, B.P. Ramos, C.D. Paspalas, Y. Shu, A. Simen, et al, (2007) alpha2A-Adrenoreceptor strengthen working memory networks by inhibiting cAMP-HCN channel signaling in prefrontal cortex; Cell (129); 397-410.
4) A. Scholten, M.K. Poh, T.A.B. van Veen, M.A. Vos, A.J.R. Heck, (2006) Analysis of the cGMP/cAMP interactome using a chemical proteomics approach in mammalian heart tissue validates sphingosine kinase type 1-interacting protein as a genuine and highly abundant AKAP; J. Proteome Res. (5); 1435-1447.