Mempro™ Cell-Based Acyl-CoA-binding Protein Production

Creative Biostructure has established a comprehensive and advanced membrane protein production platform based on years of experience, our scientists can provide custom Mempro™ transmembrane protein Acyl-CoA-binding protein production services using cell-based expression system.

Mempro™ cell-based protein production system is one of the most applied and advanced method to perform membrane proteins production. Acyl-CoA-binding protein (ACBP), also known as diazepam binding inhibitor and endozepine, is a relative small protein (10 KDa) that mainly binds Acyl-CoA esters. The affinity of this binding is extremely high. ACBP functions involve of carrying intracellular Acyl-CoA esters and modulating GABA receptor. ACBP consists of about 90 amino acids and shows a highly conservation through various species. ACBP is a unique protein, though Acyl-CoA-binding (ACB3) domains are identified in other functional proteins in various eukaryotic species. The ACB domain comprises four α-helices in a bowl shape conformation, exposing the Acyl-CoA-binding site. There are several conserved positive charges in ACBP, which bind the ligand through specific interactions. In mammalian cells, experiments indicated that ACBP knockdown leads to growth arrest and cell death.

Figure 1. Functions of ACBP. (BBA-Molecular and Cell Biology of Lipids, 2014)

Creative Biostructure can provide high-quality membrane protein production services, and we can apply various Mempro™ cell-based protein production systems including:

•      Mempro™ Acyl-CoA-binding Protein Production using Mammalian Cells System
Creative Biostructure can offer a great serviece-Mempro™ protein production in mammalian cells system. This system helps to correct folding and post-translational modifications for membrane protein. We have proprietary Mempro™ protein production in mammalian cells system with the best structural and functional features of target eukaryotic membrane proteins from numerous mammalian cell types.

•      Mempro™ Acyl-CoA-binding Protein Production using Bacterial Cells System
Escherichia coli (E. coli) is the most widely used bacterial host for membrane protein production. Lemo21 (DE3) strain is optimized for our Mempro™ protein production in our bacterial cells system. Whole-cell fluorescence can be used to monitor the membrane protein-GFP fusions production by Lemo21 (DE3), which enables the integrity of the production. The fluorescence assay also facilitates the choice of an optimally detergent to solubilize the target from the membrane, and to monitor the stability of this target membrane protein within this detergent.

•      Mempro™ Acyl-CoA-binding Protein Production using Insect Cells System
The most applied system for eukaryotic membrane protein production is in cultured insect cells and larvae. Mempro™ protein production using insect cells system can increase the expression level and reduce the truncated proteins compared to E. coli system according to similar codon usage rules. Our innovational insect cells system based on baculovirus is advanced in several aspects as below:
   •   Accuracy;
   •   Easy scale-up;
   •   High membrane protein yield;
   •   Proper folding and post translation modifications;
   •   Use of cell lines ideal for suspension culture.

•      Mempro™ Acyl-CoA-binding Protein Production using Yeast Cells System
Single-celled yeast is an easy, quick and economic culture host and able to apply multiple post translation modifications for eukaryotic membrane protein. Creative Biostructure has developed several strategies to improve yields per cell through optimizing the expression plasmid, host cell and culture conditions.

Creative Biostructure provides Mempro™ cell-based protein production services from expressing, isolating, purifying to crystallizing the membrane protein Acyl-CoA-binding protein.

We provide an array of Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
J. A. Aznar-Moreno, et. al. (2016). Characterization of a small acyl-CoA-binding protein (ACBP) from Helianthus annuus L. and its binding affinities. Plant Physiology and Biochemistry, 102, 141-150.
N. H. Raboanatahiry, et al. (2015). Computational Prediction of acyl-coA Binding Proteins Structure in Brassica napus. Plos One, 10(6): e0129650.
M. Bloksgaard, (2013). Acyl-CoA binding protein and epidermal barrier function. BBA-Molecular and Cell Biology of Lipids, 1841(3), 369–376.