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Stabilization of Human Multidrug Resistance Protein 4 (MRP4/ABCC4) Using Novel Solubilization Agents
- Source :
- Slas Discovery
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- Membrane proteins (MPs) are important drug discovery targets for a wide range of diseases. However, elucidating the structure and function of native MP is notoriously challenging as their original structure has to be maintained once removed from the lipid bilayer. Conventionally, detergents have been used to solubilize MP with varying degrees of success concerning MP stability. To try to address this, new, more stabilizing agents have been developed, such as calixarene-based detergents and styrene–maleic acid (SMA) copolymer. Calixarene-based detergents exhibit enhanced solubilizing and stabilizing properties compared with conventional detergents, whereas SMA is able to extract MPs with their surrounding lipids, forming a nanodisc structure. Here we report a comparative study using classical detergents, calixarene-based detergents, and SMA to assess the solubilization and stabilization of the human ABC transporter MRP4 (multidrug resistance protein 4/ABCC4). We show that both SMA and calixarene-based detergents have a higher solubility efficiency (at least 80%) than conventional detergents, and show striking overstabilization features of MRP4 (up to 70 °C) with at least 30 °C stability improvement in comparison with the best conventional detergents. These solubilizing agents were successfully used to purify aggregate-free, homogenous and stable MRP4, with sevenfold higher yield for C4C7 calixarene detergent in comparison with SMA. This work paves the way to MRP4 structural and functional investigations and illustrates once more the high value of using calixarene-based detergent or SMA as versatile and efficient tools to study MP, and eventually enable drug discovery of challenging and highly druggable targets.
- Subjects :
- 0301 basic medicine
Druggability
ATP-binding cassette transporter
02 engineering and technology
Biochemistry
Analytical Chemistry
MRP4/ABCC4
03 medical and health sciences
Calixarene
Humans
calixarene detergent
membrane protein
SMA
Solubility
Lipid bilayer
Nanodisc
Original Research
Protein Stability
Chemistry
021001 nanoscience & nanotechnology
Combinatorial chemistry
Recombinant Proteins
stabilization
030104 developmental biology
Membrane protein
Thermodynamics
Molecular Medicine
Multidrug Resistance-Associated Proteins
0210 nano-technology
Biotechnology
Subjects
Details
- ISSN :
- 24725552
- Volume :
- 24
- Database :
- OpenAIRE
- Journal :
- SLAS Discovery
- Accession number :
- edsair.doi.dedup.....949723a5008c3b0455a2b90c757f02f5
- Full Text :
- https://doi.org/10.1177/2472555219867074