Membrane Protein
Structure
Determination
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Cryo-EM for Membrane Protein Structure: Comprehensive Service by Creative Biostructure, Slides of Biology
Membrane proteins are essential for various cellular functions, but their structural understanding lags behind that of soluble proteins due to challenges in overexpression, solubilization, purification, and crystallization. Creative Biostructure offers a membrane protein pipeline using Cryo-Electron Microscopy (cryo-EM) to determine membrane protein structures, even for intractable targets. Their advanced electron microscopes, powerful software, and expertise in membrane protein production enable high-resolution imaging and three-dimensional reconstruction. This service provides several advantages, including compatibility with various detergent methods, rapid visualization, single-particle reconstruction, and a preferred method for intractable protein targets.
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2020/2021
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Membrane Protein
Structure
Determination
Membrane proteins are ubiquitous in biology and they play important roles
in cells as receptors, enzymes, ion channels, and transporters. Despite this,
our structural understanding has lagged behind that of their soluble
counterparts. The first challenge lies in their overexpression, solubilization,
purification and reconstitution into membrane mimetics, which often result
in a low protein yield. While X-ray crystallography has proven to be a
powerful tool over time for the study of protein structures, structure
determination of membrane proteins remains a big challenge even with well
solubilized and purified protein, due to the difficulties in forming well-
diffracting protein crystals. Many membrane proteins have proven to be
intractable to crystallization despite intense efforts. To address these
challenges, cryo-Electron Microscopy (cryo-EM)represents an attractive
alternative approach to studying the structural biology of membrane
proteins.
The advantages of our service include:
State-of-the-art electron microscopes with powerful software for data
processing
Only small amount of protein sample required (mg scale) for EM studies
Compatible with most kinds of detergents and other membrane protein
solubilization methods
Rapid visualization of large membrane proteins using negative staining TEM
Sample preparation by cryo-fixation to capture their near-native
conformational states
Single-particle reconstruction to build 3D structural models of membrane
proteins at high resolution
A preferred method for protein targets that are intractable to crystallization