Creative Biostructure has established an excellent membrane protein production platform after years of experience. Our custom Mempro™ major facilitator superfamily production service is based on our unique cell-based expression system.
Mempro™ cell-based protein production system is one of our most adopted and advanced systems for membrane proteins production. Major facilitator superfamily (MFS) transporters enable to transport various small compounds through biological membranes. MFS transporters are existed in all branches of life from human to bacteria. According to the transport mode, MFS transporters can be characterized into three groups: uniporters, which transport a single substrate; symporters, which transport a substrate associated with a coupling ion; and antiporters transport. Human major facilitator superfamily carries out various functions between cells, including nutrient absorption and renal, hepatic clearance, transportation of nutrients, metabolites, signaling molecules. Major facilitator superfamily transporter dramatically performs conformational change during the transportation cycle, the surrounding lipids are assumed to have influence on the function of the transporter. The systematically exploration of interaction between specific lipids and MFS transporters is in crucial requirement of native MSF production in native environment.
Figure 1 A schematic of the MSF transpoter clamp-and-switch model. (Nature Reviews Molecular Cell Biology, 2016)
Creative Biostructure can offer the best membrane protein production services, and we can apply various Mempro™ cell-based protein production systems including:
• Mempro™ Major Facilitator Superfamily Production in 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 to choose an optimally detergent to solubilize the target from the membrane, and to monitor the stability of this target membrane protein in detergent environment.
• Mempro™ Major Facilitator Superfamily Production in Yeast Cells System
The advantage of single-celled yeast as the membrane protein production culture host is easy, quick and economic. Single-celled yeast is able to apply multiple post translation modifications for eukaryotic membrane protein. The most widely used two species of yeast for membrane protein production are Pichia pastoris (P. pastoris) and Saccharomyces cerevisiae (S. cerevisiae). Creative Biostructure has developed several strategies to improve yields per cell through optimizing the expression plasmid, host cell and culture conditions.
• Mempro™ Major Facilitator Superfamily Production in Insect Cells System
The most used system in eukaryotic membrane protein production is insect cells and larvae system. Mempro™ protein production using insect cells system can not only increase the expression level but also 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:
• Easy scale-up;
• High membrane protein yield;
• Use of cell lines ideal for suspension culture;
• Proper folding and post translation modifications.
• Mempro™ Major Facilitator Superfamily Production in Mammalian Cells System
Creative Biostructure can provide another great service as known as Mempro™ protein production in mammalian cells system. Mammalian cells 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 including human embryonic kidney cells (HEK-293), Chinese hamster ovary cells (CHO), monkey kidney fibroblast cells (COS-7) and baby hamster kidney cells (BHK-21).
Creative Biostructure provides Mempro™ cell-based protein production services from expressing, isolating, purifying to crystallizing the major facilitator superfamily transporters.
We offer an array of Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
E. M. Quistgaard, et. al. (2016). Understanding transport by the major facilitator superfamily (MFS): structures pave the way. Nature Reviews Molecular Cell Biology, 17, 123-132.
Y. Nieng, et al. (2015). Structural Biology of the Major Facilitator Superfamily Transporters. Annual Review of Biophysics, 44: 257–283.
A. K. Redhu, (2016). MFS transporters of Candida species and their role in clinical drug resistance. FEMS Yeast Research, 16(4), fow043.