Membrane proteins play crucial roles in cellular function regulation and signal transduction and hold immense value in biological research and drug development [1]. However, traditional expression systems struggle to achieve efficient expression and extraction of membrane proteins due to their complex spatial structures and hydrophobic characteristics [2]. The detergent micelle technology platform forms detergent micelle clusters on the cell membrane, simulating the cellular membrane environment, enabling the expression and extraction of membrane proteins [3].
Detergent micelle technology can be applied to various expression systems, including bacteria, yeast, insect cells, mammalian cells, and cell-free expression systems. Each expression system has its unique advantages and disadvantages, requiring selection based on actual needs.
The advantage of the cell-free expression system lies in obtaining target proteins in a shorter time without the need for cell culture and separation steps, while providing better control over experimental conditions [4]. By applying detergent micelle technology, the cell-free expression system can achieve expression and stabilization of membrane proteins by adding suitable detergents [5]. However, cell-free expression systems may have limitations in protein translation, folding, and modification [6].
A critical step in extracting and purifying membrane proteins using detergent micelle technology is selecting the appropriate detergent. The type, concentration, and temperature of the detergent have significant effects on the extraction efficiency of membrane proteins [7].
During the experimental procedure, it is necessary to try different detergents to optimize protein extraction. Furthermore, the choice of purification methods also has a significant impact on the quality of the final membrane proteins, such as using techniques like affinity chromatography, ion exchange chromatography, and gel filtration chromatography [8].
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Based on CUSABIO's long-term experience in membrane protein expression and purification, the detergent platform technology effectively improves the content of target membrane proteins in total proteins while ensuring their correct conformation and stability. The platform's advantages are as follows:
CUSABIO has successfully developed scarce products such as 12-transmembrane SLC7A11 proteins, 7-transmembrane EDNRB, ACKR1, CCR2 proteins, 6-transmembrane AQPZ, AQP1 proteins, 3-transmembrane SLC31A1 proteins, and 1-transmembrane SLFN12L proteins, through its improved membrane protein purification technology.
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Three Technologies Help You Obtain Full-Length Muti-Transmembrane Proteins
References
[1] Overington, J. P., Al-Lazikani, B., & Hopkins, A. L. (2006). How many drug targets are there? Nature Reviews Drug Discovery, 5(12), 993-996.
[2] Bill, R. M. (2015). Playing catch with membrane proteins. Science, 347(6226), 1072-1073.
[3] Zhang, M., & Chiu, C. (2019). Membrane protein structure determination using detergent micelle platforms. FEBS Journal, 286(2), 418-428.
[4] Smith, M. T., Hawes, A. K., & Bundy, B. C. (2013). Reengineering viruses and virus-like particles through chemical functionalization strategies. Current Opinion in Biotechnology, 24(4), 620-626.
[5] Shimizu, Y., Inoue, A., Tomari, Y., Suzuki, T., Yokogawa, T., Nishikawa, K., & Ueda, T. (2001). Cell-free translation reconstituted with purified components. Nature Biotechnology, 19(8), 751-755.
[6] Carlson, E. D., Gan, R., Hodgman, C. E., & Jewett, M. C. (2012). Cell-free protein synthesis: applications come of age. Biotechnology Advances, 30(5), 1185-1194.
[7] Seddon, A. M., Curnow, P., & Booth, P. J. (2004). Membrane proteins, lipids and detergents: not just a soap opera. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1666(1-2), 105-117.
[8] Grisshammer, R., & Tate, C. G. (1995). Overexpression of integral membrane proteins for structural studies. Quarterly Reviews of Biophysics, 28(3), 315-422.