Related Experiment Video

Updated: Oct 2, 2026

Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration
10:43

Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration

Published on: May 19, 2016

PKC epsilon is associated with myosin IIA and actin in fibroblasts

Karen England1, David Ashford, Daniel Kidd

  • 1Department of Biology, University of York, York YO10 5DD, UK. k.england@ucc.ie

Cellular Signalling
|March 19, 2002
PubMed

Insights

Protein kinase C (PKC) epsilon interacts with myosin IIA and actin in fibroblast stress fibers. This PKC epsilon-actin-myosin complex is crucial for cell spreading after passage.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Protein kinase C (PKC) epsilon is involved in various cellular processes.
  • The molecular interactions of PKC epsilon in fibroblasts are not fully understood.

Purpose of the Study:

  • To identify proteins that coimmunoprecipitate with PKC epsilon in fibroblasts.
  • To investigate the role of PKC epsilon in fibroblast cell spreading.

Main Methods:

  • Matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS) was used to identify coimmunoprecipitating proteins.
  • Immunofluorescence microscopy was employed to visualize the localization of PKC epsilon, actin, and myosin.
  • Pharmacological inhibitors were used to assess the functional role of PKC and myosin in cell spreading.

Main Results:

  • Myosin IIA was identified as a novel binding partner of PKC epsilon, alongside known interactors actin, beta'Cop, and cytokeratin.
  • PKC epsilon was found to colocalize with actin and myosin in actomyosin stress fibers within fibroblasts.
  • Inhibition of PKC, myosin ATPase activity, or microfilaments impaired fibroblast spreading.

Conclusions:

  • PKC epsilon forms a complex with actin and myosin IIA within actomyosin stress fibers.
  • This PKC epsilon-actin-myosin complex plays a significant role in fibroblast cell spreading.

More Related Videos

Aip1p Dynamics Are Altered by the R256H Mutation in Actin08:57

Aip1p Dynamics Are Altered by the R256H Mutation in Actin

Published on: July 30, 2014

Assaying Protein Kinase Activity with Radiolabeled ATP08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

Related Concept Videos

Actin Polymerization and Cell Motility01:13

Actin Polymerization and Cell Motility

Actin is a family of globular proteins that are highly abundant in eukaryotic cells. It makes up approximately 1-5% of total cell protein concentration. Actin monomers polymerize to form a complex network of polarized filaments, the actin cytoskeleton, that plays a crucial role in many cellular processes, including cell motility, division, endocytosis, and metastasis of cancer cells.
Actin cytoskeleton dynamics can produce pushing, pulling, and resistance forces that help the cell to migrate.
Overview of Myosin Structure and Function01:15

Overview of Myosin Structure and Function

Myosins are a family of molecular motor proteins, first identified in the skeletal muscles, where they are responsible for muscle contraction. Along with their role in muscle contraction, these proteins also play a role in the intracellular transport of molecules and vesicles. There are twenty-four classes of myosins based on their domain sequence and organization. Of the twenty-four, six classes (Myosin I, Myosin II, Myosin V, Myosin VI, Myosin VII, and Myosin X)  have been well characterized.
Cytoskeletal Accessory Proteins01:13

Cytoskeletal Accessory Proteins

The cytoskeleton is an essential cell component that plays several structural and functional roles. However, the filaments that make up the cytoskeleton cannot function independently and depend on the accessory or ancillary proteins to effectively carry out their function. Accessory proteins associate with cytoskeletal filaments and their monomers, aiding filament formation and function. They also help in the cross-communication among cytoskeletal filaments. Cytoskeletal accessory proteins are...
The Role of Actin and Myosin in Non-muscle Cells01:10

The Role of Actin and Myosin in Non-muscle Cells

Actin and myosin or actomyosin filaments also play a significant role in cells other than those involved in muscle contraction (which occurs within the sarcomere of muscle cells). The mechanism of non-muscle cell contractile bundles was first observed in Dictyostelium and Acanthamoeba. In non-muscle cells, two bundles are commonly found: stress fibers and actomyosin adherence belts. These contractile bundles are smaller and less organized than the ones found in muscle cells. They  are held...
Introduction to Actin01:26

Introduction to Actin

Actin is a highly conserved cytoskeletal protein found abundantly in eukaryotic cells. It constitutes 10% weight of the total cellular protein in muscle cells, while in non-muscle cells, it is lower and makes up around 1–5 percent of the total cell protein. Actin found in the unicellular amoebae and complex multicellular animals is around 80% similar, demonstrating their conservation over a billion years of evolution.  Actin coding genes are conserved within species and across different species.
Actin Filament Depolymerization01:19

Actin Filament Depolymerization

Actin filaments (F-actin) are composed of actin subunits. The dissociation of actin monomers can occur from either end of F-actin. The rate of dissociation is faster from the minus-end or the pointed end, where the actin subunits exist with a bound ADP, together known as ADP-actin. The depolymerization of F-actin is aided by proteins, including the actin-depolymerizing factor (ADF) and cofilin family of proteins, gelsolin, and glia maturation factor (GMF).
In F-actin, the ADF/cofilin proteins...