Related Experiment Video

Updated: May 22, 2026

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
07:17

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex

Published on: August 2, 2021

Regulation of protein kinase C inactivation by Fas-associated protein with death domain

Wei Cheng1, Lu Wang, Rong Zhang

  • 1State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210093, China.

The Journal of Biological Chemistry
|May 15, 2012
PubMed

Insights

Fas-associated protein with death domain (FADD) regulates conventional protein kinase C (cPKC) dephosphorylation and stability. FADD deficiency enhances cPKC activity, promoting cell motility and chemotaxis in non-apoptotic processes.

Area of Science:

  • Cellular Biology
  • Molecular Signaling
  • Biochemistry

Background:

  • Protein kinase C (PKC) is crucial for cellular functions.
  • Fas-associated protein with death domain (FADD) regulates apoptosis and non-apoptotic processes.
  • The interaction between PKC and FADD in non-apoptotic signaling remains unclear.

Purpose of the Study:

  • To investigate the role of FADD in the regulation of conventional PKC (cPKC) in non-apoptotic cellular processes.
  • To elucidate the mechanism by which FADD influences cPKC activity and stability.

Main Methods:

  • Phosphorylation site mutation analysis of FADD (S191D).
  • Co-immunoprecipitation assays to study FADD-PP2A interaction.
  • Western blotting to assess cPKC phosphorylation, stability, and activity.
  • Cell-based assays for cytoskeleton reorganization, motility, and chemotaxis.

Main Results:

  • FADD is essential for maintaining the dephosphorylation of activated cPKC.
  • Phosphorylation of FADD at Ser-191 negatively regulates its function.
  • FADD interacts with protein phosphatase 2A (PP2A), mediating cPKC dephosphorylation.
  • FADD deficiency results in increased cPKC stability and activity, leading to enhanced cytoskeleton reorganization, cell motility, and chemotaxis.

Conclusions:

  • FADD plays a novel non-apoptotic role in modulating cPKC dephosphorylation, stability, and signaling termination.
  • FADD acts as a crucial regulator of cPKC activity through interaction with PP2A.
  • Dysregulation of FADD impacts cellular processes like motility and chemotaxis via altered cPKC signaling.

More Related Videos

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes09:40

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

Published on: September 28, 2018

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry05:29

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Published on: March 24, 2023

Related Concept Videos

The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
The Extrinsic Apoptotic Pathway01:17

The Extrinsic Apoptotic Pathway

The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a rapamycin-insensitive companion...
Regulation of the Unfolded Protein Response01:31

Regulation of the Unfolded Protein Response

Inositol-requiring kinase one or IRE1 is the most conserved eukaryotic unfolded protein response (UPR) receptor. It is a type I transmembrane protein kinase receptor with a distinctive site-specific RNase activity. As the binding mechanics of the misfolded proteins with the N-terminal domain of IRE-1 are unclear, three binding models — direct, indirect, and allosteric -- are proposed for receptor activation. Nevertheless, it is known that once a misfolded protein associates with IRE1, it...
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...