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Related Concept Videos

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.
Formation of Higher-order Actin Filaments01:11

Formation of Higher-order Actin Filaments

The polymerization of G-actin monomers into filamentous F-actin is a multi-step process. Once the F-actins are formed, they can bundle together in different arrangements to form higher-order networks and regulate cellular functions. Common examples include the formation of lamellipodia and filopodia at the cell's leading edge by actin reorganization in a migrating cell. The microvilli on the brush border epithelial cells are also formed through the F-actin network.
The high-order actin networks...
Generation of Straight or Branched Actin Filaments01:14

Generation of Straight or Branched Actin Filaments

The straight or branched structure formation of actin filaments is controlled by nucleating proteins such as the formins and Arp2/3 complex. Formin-mediated assembly results in straight filaments, whereas Arp2/3 protein complex-mediated assembly results in branched actin filaments.
Arp2/3 Complex
Arp2/3 complex is a seven-subunit complex consisting of two proteins similar to actin- Arp2 and Arp3, and five other subunits that help keep Arp2 and Arp3 inactive. When required, the complex is...
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.
Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
Assembly of Cytoskeletal Filaments01:18

Assembly of Cytoskeletal Filaments

Cytoskeletal filaments are polymeric forms of smaller protein subunits. However, individual cytoskeletal filaments may easily disassemble or associate with other similar filaments to form rigid structures. Microfilaments, made of actin monomers, rely on actin-binding proteins to form bundles and create networks of individual actin filaments. Microtubules rely on microtubule-associated proteins (MAPs) to form sturdy cylindrical structures. However, the proteins involved in forming complex...

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Related Experiment Video

Updated: Jul 7, 2026

Measuring Protein Binding to F-actin by Co-sedimentation
06:17

Measuring Protein Binding to F-actin by Co-sedimentation

Published on: May 18, 2017

Synthetic actin-binding domains reveal compositional constraints for function.

Maria Lorenzi1, Mario Gimona

  • 1Consorzio Mario Negri Sud, Department of Cell Biology and Oncology, Via Nazionale 8a, I-66030 Santa Maria Imbaro, Italy.

The International Journal of Biochemistry & Cell Biology
|February 26, 2008
PubMed
Summary
This summary is machine-generated.

The arrangement of calponin homology domains is critical for actin binding. Specific type 1 and type 2 domain interactions regulate actin dynamics and specificity, independent of their position.

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Last Updated: Jul 7, 2026

Measuring Protein Binding to F-actin by Co-sedimentation
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Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin
07:53

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin

Published on: March 28, 2008

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Actin-binding domains are crucial for cellular processes.
  • The canonical type 1/type 2 arrangement of calponin homology domains is common in actin-binding proteins.
  • The precise role of calponin homology domain arrangement in actin binding and filament dynamics is not fully understood.

Purpose of the Study:

  • To investigate the functional requirements of calponin homology (CH) domain arrangement in actin-binding domains.
  • To generate and characterize synthetic actin-binding domains with altered CH domain composition and position.
  • To elucidate the specific contributions of type 1 and type 2 CH domains to actin binding and filament regulation.

Main Methods:

  • Construction of synthetic actin-binding domains using the alpha-actinin-1 actin-binding domain as a scaffold.
  • Alteration of the position and composition of calponin homology (CH) domains.
  • In vitro actin-binding assays and expression in transfected cells to assess domain function.

Main Results:

  • Single and homotypic type 2 CH domain tandems failed to bind actin.
  • Single and tandem type 1 CH domain arrays bound actin but caused defective filament turnover and aberrant bundling.
  • An inverted CH domain arrangement (type 1/type 2) functioned correctly in isolation and within the full alpha-actinin molecule.

Conclusions:

  • Actin-binding domain function requires specific interactions between type 1 and type 2 calponin homology domains.
  • Type 1 CH domains provide filament binding, while type 2 CH domains regulate binding dynamics and specificity.
  • The precise positioning of CH domains is less critical than their specific type and interaction for proper actin binding.