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Related Concept Videos

Introduction to Membrane Proteins01:16

Introduction to Membrane Proteins

81.7K
The cell membrane, or plasma membrane, is an ever-changing landscape. It is described as a fluid mosaic where various macromolecules are embedded in the phospholipid bilayer. Among the macromolecules are proteins. The protein content varies across cell types. For example, mitochondrial inner membranes contain ~76% protein content, while myelin contains ~18% protein content. Individual cells contain many types of membrane proteins—red blood cells contain over 50—and different cell...
81.7K
Membrane Proteins01:30

Membrane Proteins

30.6K
Plasma membranes have integral transmembrane proteins involved in facilitated transport. These proteins are collectively referred to as transport proteins, and they function as either channels for the material or as carriers themselves. Channel proteins have hydrophilic domains exposed to the intracellular and extracellular fluids and a hydrophilic channel through their core that provides a hydrated opening for solutes to pass through the membrane layers. Passage through the channel allows...
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Structural Protein Function01:56

Structural Protein Function

30.0K
Structural proteins are a category of proteins responsible for functions ranging from cell shape and movement to providing support to major structures such as bones, cartilage, hair, and muscles. This group includes proteins such as collagen, actin, myosin, and keratin.
Collagen, the most abundant protein in mammals, is found throughout the body. In connective tissue, such as skin, ligaments, and tendons, it provides tensile strength and elasticity.  In bones and teeth, it mineralizes to...
30.0K
Mechanical Protein Functions01:58

Mechanical Protein Functions

5.7K
Proteins perform many mechanical functions in a cell. These proteins can be classified into two general categories- proteins that generate mechanical forces and proteins that are subjected to mechanical forces. Proteins providing mechanical support to the structure of the cell, such as keratin, are subjected to mechanical force, whereas proteins involved in cell movement and transport of molecules across cell membranes, such as an ion pump, are examples of generating mechanical force. 
5.7K
Protein Diffusion in the Membrane01:24

Protein Diffusion in the Membrane

5.7K
Proteins show rotational as well as lateral diffusion across the membrane. The lateral diffusion of proteins was confirmed through the cell fusion experiment where mouse and human cells were fused, resulting in hybrid cells. When the human and mouse cells fused, the specific membrane proteins on human and mouse cells were marked with the red and green-fluorescent markers, respectively. Initially, the red and green fluorescence was located on the respective hemisphere of the cell. As time...
5.7K
Detergent Purification of Membrane Proteins01:18

Detergent Purification of Membrane Proteins

6.5K
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...
6.5K

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A new functional membrane protein microarray based on tethered phospholipid bilayers.

Meriem Chadli1, Ofelia Maniti, Christophe Marquette

  • 1Univ Lyon, Université Lyon 1, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, ICBMS, UMR CNRS 5246, 43 Bd du 11 Novembre 1918, 69622 Villeurbanne, France. agnes.girard-egrot@univ-lyon1.fr.

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Summary
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A novel membrane protein biochip utilizing peptide-tethered bilayer lipid membranes (pep-tBLMs) and micropatterning was developed. This biochip enables real-time monitoring and functional assessment of membrane proteins for drug discovery screening.

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Area of Science:

  • Biotechnology
  • Biophysics
  • Chemical Engineering

Background:

  • Membrane proteins are crucial drug targets, but their study is challenging due to difficulties in reconstitution and analysis.
  • Existing biochip technologies often struggle with stability and functional preservation of membrane proteins.
  • Development of robust and functional membrane protein mimics on solid supports is essential for high-throughput screening.

Purpose of the Study:

  • To develop a novel membrane protein biochip prototype using peptide-tethered bilayer lipid membranes (pep-tBLMs) on a solid support.
  • To integrate and assess the functionality of a G protein-coupled receptor (GPCR), CXCR4, within the developed biochip.
  • To establish a platform for real-time monitoring and ligand binding studies of membrane proteins for drug discovery.

Main Methods:

  • Fabrication of peptide-tethered bilayer lipid membranes (pep-tBLMs) via micropatterning on a gold chip using a peptide spacer (P19-4H) and Ni-chelating proteoliposomes.
  • Formation of microarrays of pep-tBLMs containing the membrane protein CXCR4 within microwells.
  • Real-time visualization and characterization of pep-tBLM formation using surface plasmon resonance imaging (SPRi).
  • Assessment of CXCR4 functionality through ligand binding studies.

Main Results:

  • Successful formation of peptide-tethered bilayer lipid membranes (pep-tBLMs) in microarrays on a gold chip.
  • Real-time monitoring of pep-tBLM formation and integration of the GPCR CXCR4 was achieved using SPRi.
  • Demonstration of functional ligand binding to the reconstituted CXCR4 within the pep-tBLM microarray.
  • Validation of the biochip's potential as a screening tool for drug discovery.

Conclusions:

  • A novel membrane protein biochip based on pep-tBLMs and micropatterning has been successfully developed.
  • The biochip allows for real-time characterization and functional assessment of membrane proteins like CXCR4.
  • This technology represents a promising advancement for drug discovery screening platforms.