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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.
Coupled with recent developments in both hardware and software, the use of EM for the study of membrane proteins has been rapidly increasing. At Creative Biostructure, our in-house membrane protein pipeline covers all stages including plasmid design, protein expression, solubilization, purification, reconstruction and structure determination. With our EM platform, we use the most advanced electron microscopes equipped with direct electron detectors, constantly improving the EM procedures and making use of available resources to provide high-resolution imaging and three-dimensional reconstruction services for our customers. By combining the expertise in membrane protein production and EM, our techniques allow the structure determination of a broad spectrum of membrane proteins, such as GPCRs, ion channels, transporters, within their native environment and in a variety of conformational states, even for targets considered intractable for crystal studies.
The advantages of our service include:
Figure 1. Examples of membrane protein structures determined by cryo-EM
Creative Biostructure promises to work closely with our customers to provide tailored EM strategies for the membrane proteins of interest. The result of this service, when combined with results from other biophysical and biochemical characterization methods, can provide detailed information that will ultimately facilitate drug design process and the engineering of membrane proteins with novel function for biotechnological applications.
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