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Formation of Intermediate Filaments

Formation of Intermediate Filaments

Intermediate filaments are cytoskeletal proteins with higher tensile strength and flexibility than microfilaments and microtubules. Unlike the other two cytoskeletal proteins, intermediate filament formation lacks the enzymatic activity to hydrolyze nucleotides like ATP and GTP to generate energy for polymerization. Therefore, the formation of intermediate filaments is multistep self-assembly. The involvement of any accessory proteins in intermediate filament formation has not yet been reported.

The Structure of Intermediate Filaments

The Structure of Intermediate Filaments

The intermediate filaments are one of three widely studied cytoskeletal filaments. They are so named as their diameter (10 nm) is in between that of microfilaments (7 nm) and the microtubules (25 nm). These filaments are highly stable and can remain intact when exposed to high salt concentrations and detergents. These filaments are responsible for providing stability and mechanical support to the cells. They also help in cell adhesion and maintaining tissue integrity.
Intermediate filaments...

Disassembly of Intermediate Filaments

Disassembly of Intermediate Filaments

Intermediate filaments (IFs) do not undergo spontaneous disassembly. Enzymes, kinases, and phosphatases add and remove phosphates from specific sites to regulate their disassembly. The IF concentration in the cytoplasm also regulates the disassembly. If the concentration crosses a threshold, it activates the protein kinases in the vicinity, allowing the phosphorylation of IFs.
Keratin proteins, found at the cell periphery near cell junctions, undergo a cycle of assembly and disassembly. In Type...

Types of Intermediate Filaments

Types of Intermediate Filaments

The intermediate filaments are an essential component of the cytoskeleton. Presently six types of intermediate filament have been identified. Type I and II are acidic and basic keratin proteins. Type III is of mesodermal origin and comprises four proteins: vimentin, desmin, glial fibrillary acidic protein (GFAP), and peripherin. Vimentin is commonly found in mesenchymal cells, desmin in muscle cells, GFAP in astrocytes, while peripherin is found in peripheral nervous system neurons (PNS). Type...

Mechanism of Filopodia Formation

Mechanism of Filopodia Formation

Filopodia are thin, actin-rich cellular protrusions that play an important role in many fundamental cellular functions. They vary in their occurrence, length, and positioning in different cell types, suggesting their diverse roles.
Their main function is to guide migrating cells during normal tissue morphogenesis or cancer metastasis by recognizing and making initial contacts with the extracellular matrix. However, they can also act as stationary cell anchors or help to establish communication...

Adaptability of Cytoskeletal Filaments

Adaptability of Cytoskeletal Filaments

The cytoskeleton is a complex dynamic structure performing varied functions based on cellular requirements. The adaptability of the individual filaments in the cytoskeleton determines their ability to perform various functions within the cell. It can undergo rapid reorganization during processes like cell division or remain stable for several hours as in the interphase. The adaptability of these filaments depends on stringent regulatory mechanisms. The microfilament and microtubules of the...

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Related Experiment Video

Updated: Jul 7, 2026

Isolation of Intermediate Filament Proteins from Multiple Mouse Tissues to Study Aging-associated Post-translational Modifications

Isolation of Intermediate Filament Proteins from Multiple Mouse Tissues to Study Aging-associated Post-translational Modifications

Published on: May 18, 2017

Isolation of intermediate filaments.

Conrad L Leung¹, Ronald K H Liem

¹Columbia University, New York, New York, USA.

Current Protocols in Cell Biology

|January 30, 2008

Summary

This summary is machine-generated.

This study details methods for purifying intermediate filament (IF) proteins, including neurofilament proteins (NFL, NFM, NFH), from biological sources and recombinant systems for cell biology research.

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Area of Science:

Cell Biology
Biochemistry
Structural Biology

Background:

Intermediate filaments (IFs) are crucial cytoskeletal components found in eukaryotic cells, composed of diverse IF proteins.
IF proteins are typically insoluble, requiring denaturing conditions for purification, followed by reassembly via dialysis.

Purpose of the Study:

To provide detailed protocols for isolating and purifying intermediate filament proteins.
To illustrate IF purification using neurofilament proteins (NFL, NFM, NFH) from bovine spinal cord.
To present an alternative method for purifying NFL from a prokaryotic expression system.

Main Methods:

Purification of neurofilament proteins (NFL, NFM, NFH) from bovine spinal cord.
Reassembly of purified IF proteins into filaments through dialysis.
Purification of NFL from a prokaryotic expression system.

Main Results:

Established protocols for isolating native and recombinant intermediate filament proteins.
Demonstrated the feasibility of reassembling purified IF proteins into functional filaments.
Highlighted the efficiency of prokaryotic expression systems for producing IF proteins.

Conclusions:

Detailed purification protocols are essential for studying intermediate filament protein structure and function.
Recombinant protein expression offers a straightforward and potentially preferred method for IF purification.
Understanding IF purification is key to advancing research in cell structure and dynamics.