The following protocols are being standardized based on comparative studies of over 50 membrane proteins which have been purified intact from Escherichia coli, Pichia pastoris, insect, mammalian cells, and animal, fly and plant tissue using the SMALP system:
- Production of activated styrene maleic acid
- We're sharing recommended SMA polymers including Xiran® SL 25010 P20, SL 30010 P20 and SL 40005 P20, simply contact us for free samples.
- Preparation of cellular membrane material for SMALP'ing
- DIBMA protocol and Sandro Keller's talk
- Reaction of activated styrene maleic acid with cellular membrane
- Purification of SMALP-encapsulated proteins
- Biophysical analysis of SMALPs by methods including:
differential scanning calorimetry
dynamic light scattering
Fourier transform infrared
nuclear magnetic spectroscopy
small angle neutron scattering
- Biochemical assays to measure ligand affinities and catalytic activities
- Proteomic and lipidomic analysis of SMALPed assemblies including the metabolon
- Structural analysis of SMALP-protein complexes with crystal and electron microscopy-based trimeric structures appearing.
Protocols are described in papers including Dafforn TR, Lee SC, Knowles TJ, Postis V, Jamshad M, Parslow R, Lin YP, Goldman A, Sridhar P, Overduin M, Muench SP.A Method for Detergent-free isolation of Membrane Protein with its Local Lipid Environment, Nature Protocols, 11, 1149-1162.
Join us for the next meeting which includes a practical workshop and open discussion of SMALP protocols
In the meantime feel free to discuss below.
SMA: styrene maleic anhydride, the co-polymer which can be activated by hydrolysis to form styrene maleic acid. Sources include Polyscope.
SMALP: styrene maleic acid lipid particle, a model membrane system which contains activated SMA and is used to solubilize native membrane proteins or lipid bilayer sections directly from biological materials including whole cells without requiring conventional detergents at any stage.
nanodisc: a synthetic model membrane system that is used for the study of membrane proteins and is composed of an inner lipid bilayer section and an outer rim of two amphipathic proteins known as membrane scaffolding proteins.
Lipodisq®: a trademarked product licensed by Malvern Cosmeceutics Limited to companies including Sigma Aldrich.