Creative Biostructure provides advanced custom Mempro™ membrane protein production services for single-helix ATPase regulators using cell-free expression system. Our patent cell-free expression system can best preserve the native conformation of desired transmembrane proteins.
Mempro™ cell-free protein production system is the innovative platform to produce membrane proteins, which can bypass the limitations that encountered in cell-based protein production system, including host cell toxicity, low yield and solubilization and purification using detergents. Single-helix ATPase regulators are a large group of bitopic proteins with alpha-helical transmembrane anchors and regulate ATPases. It is well known that alpha-helices and beta-barrels are the main components of the transmembrane domains in transmembrane proteins. Single-helix ATPase regulators can be classified two families, including calcium ATPase regulators and FXYD regulators. Calcium ATPase regulators locate in endoplasm reticulum (ER) and regulate calcium homeostasis. Members of calcium ATPase regulators family include phospholamban, sarcolipin and cytoplasmic helix. FXYD regulators locate in the plasma membrane of eukaryo, members of FXYD regulators family are tissue-specific modulators of Na, K-ATPase. FXYD1 mainly increases the apparent affinity for intracellular Na+ of Na, K-ATPase. FXYD2 and FXYD4 can oppositely regulate the apparent affinity for Na+ of Na, K-ATPase.
Figure 1. A schametic of transmembrane protein. 1. bitopic membrane protein.2. a polytopic transmembrane α-helical protein. 3. a polytopic transmembrane β-sheet protein. (Wikipedia)
Creative Biostructure has rich professional experience in high-yield single-helix ATPase regulators production using cell-free protein expression platform, we can provide various strategies for Mempro™ cell-free membrane protein production, including:
Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is popularly used for single-helix ATPase regulators production. Wheat germ extract can provide eukaryotic translation system for single-helix ATPase regulators expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for single-helix ATPase regulators expression, which is developed in the early stage that transfected insect cells in combination with vectors derived from the baculovirus. Crude rabbit reticulocyte lysate is also the most widely employed for eukaryotic cell-free transmembrane protein expression. To select an appropriate source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.
With the Mempro™ cell-free protein production platform, Creative Biostructure is capable of expressing, purifying and crystallizing single-helix ATPase regulators to facilitate the study of their biological functions.
We can provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
D. H. MacLennan and E. G. Kranias. (2003) Phospholamban: a crucial regulator of cardiac contractility. Nat. Rev. Mol. Cell Biol., 4(7): 566-77.
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
K. Geering (2005). Function of FXYD proteins, regulators of Na, K-ATPase. J. Bioenerg. Biomembr., 37(6): 387-92.