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Mechanisms of Membrane Domain Formation00:59

Mechanisms of Membrane Domain Formation

Different physical properties of lipids and proteins allow them to localize and form distinct islands or domains in the membrane. Some membrane domains are formed due to protein-protein interactions, whereas others are formed due to the presence of specific lipids such as sphingolipids and sterols—for example, large proteins, such as bacteriorhodopsin, aggregate and create distinct domains.
Another mechanism for membrane domain formation involves membrane proteins interacting with cytoskeletal...
Protein-protein Interfaces02:04

Protein-protein Interfaces

Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...
Detergent Purification of Membrane Proteins01:18

Detergent Purification of Membrane Proteins

Detergents are used to purify the integral proteins of the membrane. The hydrophobic portion of the detergent can replace membrane phospholipids while solubilizing the membrane proteins. When detergent monomers reach a specific concentration in a solution called critical micelle concentration (CMC), they form micelles. Above CMC, the concentration of the detergent monomers remains in equilibrium with the micelle. The number of detergent monomers present in the CMC varies for each detergent, and...
Lipids as Anchors01:32

Lipids as Anchors

In the plasma membrane, the lipids forming the bilayer can also act as an anchor to tether proteins to the membrane. The three main types of lipid anchors found in eukaryotes are – prenyl groups, fatty acyl groups, and glycosylphosphatidylinositol or GPI groups. Prenyl and fatty acyl groups act as anchors on the cytosolic surface of the membrane, whereas GPI anchors proteins on the extracellular side.
The carboxy-terminal of most of the prenylated proteins, such as Ras proteins, contains the...
Membrane Fluidity01:26

Membrane Fluidity

Membrane fluidity is explained by the fluid mosaic model of the cell membrane, which describes the plasma membrane structure as a mosaic of components—including phospholipids, cholesterol, proteins, and carbohydrates—that gives the membrane a fluid character.
Mosaic nature of the membrane
The mosaic characteristic of the membrane helps the plasma membrane remain fluid. The integral proteins and lipids exist as separate but loosely-attached molecules in the membrane. The membrane is a relatively...

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Related Experiment Video

Updated: May 13, 2026

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins
09:55

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

Published on: July 4, 2016

A generalized strategy for immobilizing uniformly oriented membrane proteins at solid interfaces.

Amit Vaish1, Vitalii Silin, Marlon L Walker

  • 1National Institute of Standards and Technology (NIST) Center for Neutron Research, Gaithersburg, MD 20899, USA. amit.vaish@nist.gov

Chemical Communications (Cambridge, England)
|February 26, 2013
PubMed
Summary

We developed a method using thiol self-assembly to immobilize membrane proteins on gold surfaces. This technique successfully captured histidine-tagged G protein-coupled receptors in a specific orientation with minimal non-specific binding.

More Related Videos

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
11:13

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy

Published on: August 20, 2018

Related Experiment Videos

Last Updated: May 13, 2026

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins
09:55

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

Published on: July 4, 2016

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
11:13

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy

Published on: August 20, 2018

Area of Science:

  • Biochemistry
  • Surface Science
  • Biophysics

Background:

  • Membrane proteins are crucial for cellular functions but challenging to study.
  • Immobilization of membrane proteins in a defined orientation is essential for structural and functional analysis.
  • Existing methods often suffer from non-specific binding and loss of protein activity.

Purpose of the Study:

  • To develop a novel method for orienting and immobilizing membrane proteins at interfaces.
  • To investigate the self-assembly of thiols on gold substrates for protein capture.
  • To validate the method using a model G protein-coupled receptor (GPCR).

Main Methods:

  • Utilized self-assembly of nitrilotriacetic acid (NTA)-terminated oligo(ethylene glycol) thiols on gold (Au) substrates.
  • Immobilized histidine-tagged membrane proteins via the NTA-histidine tag interaction.
  • Characterized protein immobilization and orientation using surface-based techniques.

Main Results:

  • Achieved successful immobilization of membrane proteins at the gold-thiol interface.
  • Demonstrated capture of histidine-tagged G protein-coupled membrane receptors (GPCRs) in a defined orientation.
  • Observed minimal non-specific binding of proteins to the functionalized surface.

Conclusions:

  • The developed thiol-based self-assembly method provides a robust platform for oriented immobilization of membrane proteins.
  • This approach minimizes non-specific binding, enabling more accurate studies of membrane protein structure and function.
  • The method is suitable for various histidine-tagged membrane proteins, including GPCRs.