Mempro™ Cell-Free Acyl-CoA-binding Protein (ACBP) Production

Creative Biostructure provides high-quality protein production for acyl-CoA-binding protein (ACBP) using our patent Mempro™ cell-free expression system. Compared to protein expression in living cells, cell-free system produces the protein from gene in a more efficient manner. Moreover, it eliminates the drawback of limited production when the protein is toxic or susceptible to proteolytic degradation in cell-based expression system.

Acyl-CoA-binding protein (ACBP) is a highly conserved lipid carrier protein consisting of approximately 86-102 amino acid residues with an acetylated N-terminus. It is found to bind saturated acyl-CoA esters of chain length C12-C22 with high affinity. ACBP plays a significant role in transporting intracellular acyl-CoA and regulating cellular functions, such as cell growth, medium-chain acyl-CoA-ester synthesis. From the sequence alignment, ACBP can be divided into four groups. The first group is the basic isoform found in bovine liver (l-ACBP). The second group is the testis specific isoform (t-ACBP). The third putative group is found in brain (b-ACBP). The fourth group has longer sequences up to 533 amino acids that is suggested to be membrane bound protein. L-ACBP consists of four α-helices held together, forming a hydrophobic groove on the protein surface. The head group of acyl-CoA interacts with the residues on the rim of the binding pocket, forms a lid to protect the the acyl chain from the aqueous solution. The acyl-CoA head group binds to the l-ACBP loosely and determines the specificity of the the acyl-CoA ester.

 
Figure 1. Structure of bovine acyl-CoA binding protein in tetragonal crystal form.

Creative Biostructure has a wealth of experience in high-yield ACBP production using cell-free protein expression systems, we can provide various strategies for Mempro™ cell-free protein production, including:

• Mempro™ Cell-Free Protein Production in E. coli;
• Mempro™ Cell-Free Protein Production in Wheat Germ;
• Mempro™ Cell-Free Protein Production in Rabbit Reticulocyte;
• Mempro™ Cell-Free Protein Production in Baculovirus.

 
ACBP has been successfully expressed in E. coli. The Mempro™ Cell-Free system we employed utilizes the E. coli S30 extract, which is optimal for ACBP production. E. coli S30 extract system contains the translational machinery required for both prokaryotic and eukaryotic protein. It’s widely applied to a variety of proteins for its fast and active production.

With the Mempro™ cell-free protein production platform, ACBP can be expressed, purified and crystallized with high quality in a cost-effective manner.

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

References:
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
Rasmussen J T, FÆrgeman N J, Kristiansen K, et al. Acyl-CoA-binding protein (ACBP) can mediate intermembrane acyl-CoA transport and donate acyl-CoA for β-oxidation and glycerolipid synthesis[J]. Biochemical Journal, 1994, 299(1): 165-170.
Knudsen J, Neergaard T B F, Gaigg B, et al. Role of acyl-CoA binding protein in acyl-CoA metabolism and acyl-CoA–mediated cell signaling[J]. The Journal of nutrition, 2000, 130(2): 294S-298S.