Overcoming bottlenecks in the membrane protein structural biology pipeline.
Membrane proteins account for a third of the eukaryotic proteome, but are greatly under-represented in the Protein Data Bank. Unfortunately, recent technological advances in X-ray crystallography and EM cannot account for the poor solubility and stability of membrane protein samples. A limitation of conventional detergent-based methods is that detergent molecules destabilize membrane proteins, leading to their aggregation. The use of orthologues, mutants and fusion tags has helped improve protein stability, but at the expense of not working with the sequence of interest. Novel detergents such as glucose neopentyl glycol (GNG), maltose neopentyl glycol (MNG) and calixarene-based detergents can improve protein stability without compromising their solubilizing properties. Styrene maleic acid lipid particles (SMALPs) focus on retaining the native lipid bilayer of a membrane protein during purification and biophysical analysis. Overcoming bottlenecks in the membrane protein structural biology pipeline, primarily by maintaining protein stability, will facilitate the elucidation of many more membrane protein structures in the near future.
Life & Health Sciences, Aston University, Birmingham B4 7ET, U.K. Calixar, 60 Avenue Rockefeller, 69008 Lyon, France.
David
D
Hardy
Life & Health Sciences, Aston University, Birmingham B4 7ET, U.K.
Roslyn M
RM
Bill
Calixar, 60 Avenue Rockefeller, 69008 Lyon, France a.rothnie@aston.ac.uk ajawhari@calixar.com.
Anass
A
Jawhari
Life & Health Sciences, Aston University, Birmingham B4 7ET, U.K. a.rothnie@aston.ac.uk ajawhari@calixar.com.
Alice J
AJ
Rothnie
D000818:Animals; D003902:Detergents / Q000737:chemistry; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification; D055550:Protein Stability; D040901:Proteomics; D012995:Solubility
D003902:Detergents; D008565:Membrane Proteins
calixarenes; maltose neopentyl glycol (MNG); membrane; solubilization; structural biology; styrene maleic acid lipid particle (SMALP)