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Related Concept Videos

Protein Diffusion in the Membrane01:24

Protein Diffusion in the Membrane

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...
Single-pass Transmembrane Proteins01:25

Single-pass Transmembrane Proteins

Integral membrane proteins are tightly associated with the cell membrane and play a crucial role in cell communication, signaling, adhesion, and transport of the molecules. Some integral membrane proteins are present only in the membrane monolayer. For example, the enzyme fatty acid amide hydrolase is present in the cytoplasmic side of the membrane monolayer. In contrast, another type of integral membrane protein, also known as a transmembrane protein, spans across the membrane. Transmembrane...
Introduction to Membrane Proteins01:16

Introduction to Membrane Proteins

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 types have...
Insertion of Single-pass Transmembrane Proteins in the RER01:26

Insertion of Single-pass Transmembrane Proteins in the RER

Integral membrane proteins are proteins adhered to the lipid bilayer of a cell organelle or membrane. They can be of two types: transmembrane integral proteins that span the lipid bilayer and monotopic proteins that are attached to either side of the membrane but do not pass through it.
Integral transmembrane proteins possess transmembrane and extra membrane domains. The transmembrane domains are primarily made of 20-25 hydrophobic amino acids arranged in a helical secondary confirmation. These...
Insertion of Multi-pass Transmembrane Proteins in the RER01:29

Insertion of Multi-pass Transmembrane Proteins in the RER

The rough ER membrane synthesizes, assembles, and embeds transmembrane proteins in diverse topologies. These proteins function as transporters or channels and can remain in the ER membrane or are sent to the Golgi complex, lysosome, and cell membrane.
The multipass transmembrane proteins are the type IV integral membrane proteins with multiple topogenic sequences determining their spatial arrangement in the ER membrane. Nearly all multipass proteins lack a cleavable signal sequence and use...
What are Membranes?01:24

What are Membranes?

A cell's plasma membrane demarcates the cell's borders and determines the nature of its interaction with the environment. Cells exclude certain substances, take in others, and excrete some others in controlled quantities. The plasma membrane must be flexible to allow certain cells, such as red and white blood cells, to change their shape while passing through narrow capillaries. These are the more obvious plasma membrane functions. In addition, the plasma membrane's surface carries markers that...

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Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

Membrane protein expression: no cells required.

Federico Katzen1, Todd C Peterson, Wieslaw Kudlicki

  • 1Life Technologies, 5791 Van Allen Way, Carlsbad, CA 92008, USA.

Trends in Biotechnology
|July 21, 2009
PubMed
Summary
This summary is machine-generated.

Cell-free protein expression offers a promising solution for producing difficult-to-express membrane proteins. This review highlights recent advances, particularly nanolipoprotein particles (NLPs), for efficient protein production.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Protein Engineering

Background:

  • Membrane protein studies are hindered by challenges in producing sufficient quantities of correctly folded proteins.
  • Cell-based expression often leads to low yields, toxicity, aggregation, and misfolding of membrane proteins.

Purpose of the Study:

  • To review recent advancements in cell-free protein expression for membrane proteins.
  • To highlight the potential and limitations of novel expression strategies.
  • To emphasize the application of nanolipoprotein particles (NLPs) in this domain.

Main Methods:

  • Review of recent literature on cell-free protein expression techniques.
  • Analysis of newly developed approaches for membrane protein production.
  • Focus on the utilization and efficacy of nanolipoprotein particles (NLPs).

Main Results:

  • Cell-free expression systems show significant promise for overcoming challenges in membrane protein production.
  • Nanolipoprotein particles (NLPs) are emerging as a key technology for efficient and functional membrane protein expression.
  • Various new approaches offer improved yields and protein quality compared to traditional methods.

Conclusions:

  • Cell-free protein expression, especially with nanolipoprotein particles (NLPs), is a powerful tool for advancing membrane protein research.
  • Further development of these techniques will accelerate structural and functional studies of membrane proteins.
  • The field is rapidly evolving, offering new possibilities for producing complex and challenging protein targets.