Mempro™ Detergent-Free C2 Domain-Containing Protein Production

Creative Biostructure can provide high-quality custom Mempro™ C2 domain-containing protein production services using detergent-free expression system.

C2 domain is a region containing 130 amino acids that is originally found in proteins that can bind phospholipids in a calcium-dependent or -independent manner. When calcium is bound with C2 domain, which can insert directly into the cell membranes. Proteins containing C2 domain only exist in eukaryotes. There are two types of C2 domains, including C₂A and C₂B. Take Synaptotagmin-1 as an example, the C₂B-domain of synaptotagmin-1 is a Ca²⁺-dependent phospholipid binding protein, however, Ca²⁺ binding can result in conformational changes Ca²⁺ in the C₂B-domain of synaptotagmin-1. This result can be also obtained from the C₂A-domain of synaptotagmin-1. Creative Biostructure can provide the best production of C2 domian-containing proteins to facilitate the structural and functional studies.

Figure 1. C2 domain of Synaptoagmin-1. (Neuron, 2001)

Creative Biostructure has great expertise in high-yield C2 domain-containing protein production using detergent-free membrane protein expression system, we can carry out various strategies for Mempro™ detergent-free membrane protein production, including:

• Mempro™ C2 domain-containing protein production using nanodiscs

Nanodiscs are the self-assembly nanotechnology to stabilize membrane proteins removed from the membrane by membrane scaffold proteins (MSPs) in no need of detergent. Creative Biostructure can provide combination of C2 domain-containing protein expressed by cell-free expression system into pre-assembly nanodiscs during purification process.

• Mempro™ C2 domain-containing protein production using amphipols

The amphipathic amphipols have the ability to “trap” around membrane proteins, allowing them to stay folded. Creative Biostructure can apply A8-35 to solubilize C2 domain-containing proteins.

• Mempro™ C2 domain-containing protein production using poly (styrene-co-maleic acid) lipid particles (SMALPs)

The SMALPs are self-assembled by the simple addition of the SMA co-polymer. At neutral or alkaline pH, a disc-like structure assembles itself, encapsulating C2 domain-containing proteins in a form amenable to be purified.

These novel detergent-free purification approaches for C2 domain-containing protein production can be obtained easily, and facilitate more comprehensively structural and functional analysis.

Creative Biostructure can provide various custom Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
C2 domain. (https://en.wikipedia.org/wiki/C2_domain)
C. Tribet, et al. (1996). Amphipols: Polymers that keep membrane proteins soluble in aqueous solutions. Proc Natl Acad Sci U. S. A. 93: 15047-15050.
I. Fernandez, et al. (2001). Three-dimensional structure of the synaptotagmin 1 C2B-domain: synaptotagmin 1 as a phospholipid binding machine. Neuron, 32(6): 1057-1069.
M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans., 39: 813-818.
R. B. Sutton, et al. (1995). Structure of the first C2 domain of synaptotagmin I: a novel Ca²⁺/phospholipid-binding fold. Cell, 80(6): 929-938.
R.O. Ryan, (2010). Nanobiotechnology applications of reconstituted high density lipoprotein. J. Nanobiotechnol., 8(1):28-37.