With the most advancd technology and years of experiences in protein engineering field, Creative Biostructure can provide custom Mempro™ C1 domain-containing protein production services using cell-based expression system.
Mempro™ cell-based protein production system is commonly used for producing integral membrane proteins, such as G-protein coupled receptors (GPCRs) and ion channels. Protein kinase C conserved region 1 (C1) domains, also termed as phorbol esters/diacylglycerol binding domains, can bind the lipid second messenger diacylglycerol (DAG), as well as the phorbol esters (analogues of DAG). It is well known that C1 domain is a cysteine-rich domain which present in numerous signaling family members, including chimaerins, RasGRPs, PKDs, diacylglycerol kinases (DGKs) and so on. C1 domain consists of around 50 amino acids and is involved in the recruitment of proteins to the membrane.
Figure 1. The structural model of PKC-delta C1 domain. (Wikipedia)
Creative Biostructure has strong expertise in high-yield C1 domain-containing protein production based on cell-based protein expression system, we can provide various strategies for Mempro™ cell-based protein production, including:
There are two strategies can be performed in bacterial cells expression system, protein expressed as soluble membrane proteins or as inclusion bodies representing in cytoplasmic aggregates. Yeast cells (mostly saccharomyces cerevisiae and pichia pastoris) system, combines both prokaryotic as well as eukaryotic characteristic advantages such as single cells, fast growth rates, inexpensive media, high cell densities. Insect cells (such as Sf9, Sf21 and High Five) are also a major system for C1 domain-containing protein expression, which are developed from transfected insect cells in combination with vectors derived from the baculovirus species AcMNPV. Mammalian cells which provide near-native cell environment is another eukaryotic system for C1 domain-containing protein expression. Cell lines derived from COS, CHO, BHK-21, HEK293, HeLa and GH3 are widely used for integral membrane protein expression.
With the Mempro™ cell-based protein production platform, Creative Biostructure is capable of expressing, isolating, purifying and crystallizing C1 domain-containing proteins to facilitate the study of their biological functions.
We provide other 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. Colón-González and M. G. Kazanietz (2006). C1 domains exposed: from diacylglycerol binding to protein-protein interactions. Biochim. Biophys. Acta., 1761(8): 827-837.F. Junge, et al. (2008) Large-scale production of functional membrane proteins. Cellular Mol. Life Sci., 65 (11): 1729-1755. J. Das and G. M. Rahman (2014). C1 domains: structure and ligand-binding properties. Chem. Rev., 114 (24): 12108-12131. S. Schlegel, et al. (2013). Bacterial-based membrane protein production. Biochim. Biophys. Acta., 1843(8): 1739-1749.