AI Article Synopsis
- To study PKA activity in living cells, researchers created a fluorescent substrate linked to part of the PKA regulatory domain that has an autophosphorylation site.
- When intracellular cAMP levels rise, this substrate gets phosphorylated, leading to a reduction in its fluorescence intensity.
- In NG108-15 cells, PKA activity was primarily found in the cytosol near the nucleus, and in hippocampal neurons, L-glutamate triggers PKA activation linked to higher cAMP levels.
Article Abstract
In order to visualize the activity of the cAMP-dependent protein kinase (PKA) in living cells, we have constructed a new fluorescence PKA substrate by conjugating a fluorescence probe to a partial amino acid sequence of PKA regulatory domain II which contains a specific autophosphorylation site. The fluorescent peptide was cell-permeable and became phosphorylated when the intracellular cAMP concentration was increased, resulting in a decrease in its fluorescence intensity. In NG108-15 cells, PKA activity was localized to the cytosol around the nucleus. In cultured hippocampal neurons, addition of L-glutamate caused PKA activation associated with increase of the cellular cAMP.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/s0014-5793(97)00970-8 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
PDE4 inhibitor rolipram represses hedgehog signaling via ubiquitin-mediated proteolysis of GLI transcription factors to regress breast cancer.
J Biol Chem
January 2025
Cell and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani, Nadia, West Bengal, India, 741235. Electronic address:
Aberrant activation of the hedgehog (Hh) signaling pathway positively correlates with progression, invasion and metastasis of several cancers, including breast cancer. Although numerous inhibitors of the Hh signaling pathway are available, several oncogenic mutations of key components of the pathway, including Smoothened (Smo), have limited their capability to be developed as putative anti-cancer drugs. In this study, we have modulated the Hh signaling pathway in breast cancer using a specific FDA-approved phosphodiesterase 4 (PDE4) inhibitor rolipram.
View Article and Find Full Text PDFCharacterization of multiple binding sites on microtubule associated protein 2c recognized by dimeric and monomeric 14-3-3ζ.
FEBS J
January 2025
Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Microtubule associated protein 2 (MAP2) interacts with the regulatory protein 14-3-3ζ in a cAMP-dependent protein kinase (PKA) phosphorylation dependent manner. Using selective phosphorylation, calorimetry, nuclear magnetic resonance, chemical crosslinking, and X-ray crystallography, we characterized interactions of 14-3-3ζ with various binding regions of MAP2c. Although PKA phosphorylation increases the affinity of MAP2c for 14-3-3ζ in the proline rich region and C-terminal domain, unphosphorylated MAP2c also binds the dimeric 14-3-3ζ via its microtubule binding domain and variable central domain.
View Article and Find Full Text PDFModeling ocular surface ion and water transport by generation of lipid- and mucin-producing human Meibomian gland and conjunctival epithelial cells.
Am J Physiol Cell Physiol
January 2025
Laboratoire de Physiopathologie et Régulation des Transports Ioniques, Université de Poitiers, France.
Despite the importance of ocular surface in human physiology and diseases, little is known about ion channel expression, properties and regulation in ocular epithelial cells. Furthermore, human primary epithelial cells have rarely been studied in favor of rat, mouse and especially rabbit animal models. Here, we developed primary human Meibomian gland (hMGEC) and conjunctival (hConEC) epithelial cells.
View Article and Find Full Text PDFIdentification of prognostic biomarkers of sepsis and construction of ceRNA regulatory networks.
Sci Rep
January 2025
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Sepsis is a life-threatening severe organ dysfunction, and the pathogenesis remains uncertain. Increasing evidence suggests that circRNAs, mRNAs, and microRNAs can interact to jointly regulate the development of sepsis. Identifying the interaction between ceRNA regulatory networks and sepsis may contribute to our deeper understanding of the pathogenesis of sepsis, bring new insights into early recognition and treatment of sepsis.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!