Based on the comprehensive membrane protein production platform establised through years of experience, experters from Creative Biostructure can offer custom Mempro™ major instrinsic protein (MIP)/formate-nitrite transporter (FNT) superfamily production services using cell-based expression system.
Mempro™ cell-based protein production system is the most widely used for membrane proteins production. Members of MIP superfamily are transmembrane protein channels (six transmembrane helices, TM1-TM6; and two half-helices, LB and LE) that facilitate water and solute permeability across cell membranes and are found in eubacteria, archaea, fungi, plants and animals. It is reported that aquaporin (AQP) and aquaglyceroporin channels are the primary members of the MIP superfamily. AQPs are expressed in a wide range of tissues, often spatially located within a certain region of the cell. Members of FNT superfamily can function as channels monovalent anions, and which contain a structural domain termed the aquaporin helical fold. The Transport Classification Database (TCDB) show that the FNT superfamily in included in the MIP superfamily.
Figure 1. The topology diagram of an MIP channel. (Methods Enzymol., 2015)
Creative Biostructure can produce high quality MIP/FNT superfamily proteins, we can perform various strategies for Mempro™ cell-based protein production, including:
Escherichia coli (E. coli) is the powerful bacterial host for the production of MIP/FNT superfamily proteins. 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 MIP/FNT superfamily proteins 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 MIP/FNT superfamily proteins expression, which provide near-native cell environment. Cell lines derived from COS, CHO, BHK-21, HEK293, HeLa and GH3 can be commonly used for transmembrane protein expression.
With the Mempro™ cell-based protein production platform, Creative Biostructure is capable of expressing, isolating, purifying and crystallizing MIP/FNT superfamily proteins 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:F. Junge, et al. (2008) Large-scale production of functional membrane proteins. Cellular Mol. Life Sci., 65 (11): 1729-1755.J. Petschnigg, et al. (2011). Using yeast as a model to study membrane proteins. Curr. Opin. Nephrol. Hypertens., 20(4): 425-432.M. Mukherjee, et al. (2016). Functional diversity within the fnt superfamily of anion channels: phylogenetics & molecular dynamics studies. Biophysical J., 110(3): p117a.N. Yan (2015). Structural biology of the major facilitator superfamily transporters. Annu. Rev. Biophys., 44: 257-283.R. K. Verma, et al. (2015). Major intrinsic protein superfamily: channels with unique structural features and diverse selectivity filters. Methods Enzymol., 557: 485-520.S. Schlegel, et al. (2013). Bacterial-based membrane protein production. Biochim. Biophys. Acta., 1843(8): 1739-1749.