Mempro™ Cell-Based Pentameric Ligand-Gated Ion Channel Production

Creative Biostructure can provide custom Mempro™ pentameric ligand-gated ion channel production services using cell-based expression system.

Mempro™ cell-based protein production system is the most widely used for the production of membrane proteins.The pentameric ligand-gated ion channel (pLGIC), a single neurotransmitter receptor superfamily, can mediate fast synaptic communication (chemical transmission) through converting nerve signals into an electrical response. The pLGICs are a group of transmembrane ion channels that open to allow ions such as Na⁺, K⁺, Ca²⁺, or Cl⁻ to pass through the membrane, which are composed of five homologous subunits. It is shown that theses subunits consist of at least two different domains, including a transmembrane domain which forms the channel gate and the ion selective filter, and an extracellular domain which includes the ligand binding location (an allosteric binding site). In addition, pLGICs are divided into three classes: Cys-loop receptors, Ionotropic glutamate receptors and ATP-gated channels.

Figure 1. A schematic diagram of the topology of a pLGIC subunit. (Adv. Exp. Med. Biol., 2010)

Creative Biostructure can produce high-yield pLGICs, we can perform various strategies for Mempro™ cell-based protein production, including:

• Mempro™ Protein Production in Bacterial Cells System ;
• Mempro™ Protein Production in Yeast Cells System;
• Mempro™ Protein Production in Incest Cells System;
• Mempro™ Protein Production in Mammalian Cells System.

Escherichia coli (E. coli) is the powerful bacterial host for the production of pLGICs. Yeast cells (such as saccharomyces cerevisiae and pichia pastoris) system, combines prokaryotic as well as eukaryotic characteristics. It is a perfect eukaryotic host due to single cells, fast growth rates, inexpensive media, high cell densities. Insect cells (such as Sf9, Sf21 and High Five) are also the major system for pLGICs expression, which are developed from transfected insect cells in combination with vectors derived from the baculovirus species AcMNPV. Mammalian cells present another eukaryotic system for pLGICs expression, which provide near-native cell environment. Cell lines derived from COS, CHO, BHK-21, HEK293, HeLa and GH3 can be generally used for transmembrane protein expression.

With the Mempro™ cell-based protein production platform, Creative Biostructure is capable of expressing, isolating, purifying and crystallizing pLGICs to facilitate the study of their physiological functions.

We provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
C. Trometer, and P. Falson (2010). Mammalian membrane protein expression in baculovirus-infected insect cells. Methods Mol. Biol., 601: 105-117.
F. Junge, et al. (2008) Large-scale production of functional membrane proteins. Cellular Mol. Life Sci., 65 (11): 1729-1755.
J. A. Dent (2010). The evolution of pentameric ligand-gated ion channels. Adv. Exp. Med. Biol., 683: 11-23.
J. B. Corrie, et al. (2013). Gating of pentameric ligand-gated ion channels: structural insights and ambiguities. Structure, 21(8): 1271-1283.
S. Schlegel, et al. (2013). Bacterial-based membrane protein production. Biochim. Biophys. Acta., 1843(8): 1739-1749.