Related Experiment Video
Updated: Apr 20, 2026
13:16
Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
Published on: December 31, 2019
9.9K
Functional expression of plant membrane proteins in Lactococcus lactis
Sylvain Boutigny1, Emeline Sautron, Annie Frelet-Barrand
1CNRS, Laboratoire de Physiologie Cellulaire & Végétale, UMR 5168, 17 rue des Martyrs, 38054, Grenoble, France.
Methods in Molecular Biology (Clifton, N.J.)
|December 3, 2014
Summary
This study provides protocols for expressing and solubilizing plant membrane proteins using Lactococcus lactis. This method aids in the biochemical characterization of these challenging hydrophobic proteins.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Membrane protein study is difficult due to hydrophobicity and low abundance.
- Lactococcus lactis is a versatile host for heterologous protein production.
- This bacterium is suitable for producing prokaryotic and eukaryotic membrane proteins.
Purpose of the Study:
- To provide protocols for expressing plant membrane proteins in Lactococcus lactis.
- To detail methods for solubilizing these proteins from bacterial membranes.
- To facilitate further purification and biochemical characterization.
Main Methods:
- Utilizing Lactococcus lactis as an expression system.
- Developing protocols for protein expression.
- Implementing solubilization techniques for membrane proteins.
Main Results:
- Successful expression of plant peripheral and intrinsic membrane proteins.
- Established protocols for protein solubilization from Lactococcus membranes.
- Foundation laid for purification and characterization.
Conclusions:
- Lactococcus lactis is an effective system for producing and preparing plant membrane proteins.
- The provided protocols simplify the study of challenging membrane proteins.
- Enables detailed biochemical characterization of plant membrane proteins.
Related Concept Videos
02:09Operons
57.1K
Prokaryotes can control gene expression through operons—DNA sequences consisting of regulatory elements and clustered, functionally related protein-coding genes. Operons use a single promoter sequence to initiate transcription of a gene cluster (i.e., a group of structural genes) into a single mRNA molecule. The terminator sequence ends transcription. An operator sequence, located between the promoter and structural genes, prohibits the operon’s transcriptional activity if bound by...
57.1K
01:26Bacterial Translocation and Protein Secretion
1.1K
Bacterial protein secretion involves translocation systems to ensure proteins reach their designated locations, including the plasma membrane, periplasm, outer membrane, or the external environment. These translocation systems are vital for bacterial physiology, supporting processes like membrane assembly, enzymatic activity in the periplasm, and interactions with the external environment. The division of labor between Sec and Tat pathways ensures efficiency in handling proteins with diverse...
1.1K
01:25Inducible Operons: lac Operon
3.1K
The lac operon in Escherichia coli is a model for understanding inducible gene regulation and metabolic flexibility. It integrates local control by lactose and global regulation through catabolite repression, enabling E. coli to preferentially metabolize glucose when available and switch to lactose utilization when glucose is scarce.Structure and Function of the lac OperonThe lac operon contains three structural genes: lacZ (β-galactosidase), lacY (lactose permease), and lacA...
3.1K
01:26Insertion of Single-pass Transmembrane Proteins in the RER
19.0K
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...
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...
19.0K
01:17Gram-negative Bacterial Protein Secretion Systems
1.6K
Gram-negative bacteria utilize sophisticated protein secretion systems to transport proteins across their double-membrane envelope into the extracellular environment or host cells. Based on their mechanism of action, these systems are classified into one-step and two-step pathways.One-Step Secretion Systems (Types I, III, IV, and VI)One-step secretion systems bypass the periplasm entirely, forming a continuous channel that spans both the inner and outer membranes:Type I Secretion System (T1SS):...
1.6K
01:19Formation of Lipopolysaccharides
1.1K
Lipopolysaccharides (LPS) are crucial components of the outer membrane of Gram-negative bacteria, serving both structural and functional roles. It contributes to membrane stability and protects bacteria from host immune responses. LPS is composed of three major regions—lipid A, a core oligosaccharide, and an O antigen. The biosynthesis and assembly of LPS involve a highly coordinated set of enzymatic reactions and transport mechanisms. Additionally, LPS is recognized as an endotoxin,...
1.1K