Reconstitution of Detergent-Solubilized Membrane Proteins into Proteoliposomes and Nanodiscs for Functional and Structural Studies. 2021

Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.

Reconstitution of detergent-solubilized membrane proteins into phospholipid bilayers allows for functional and structural studies under close-to-native conditions that greatly support protein stability and function. Here we outline the detailed steps for membrane protein reconstitution to result in proteoliposomes and nanodiscs. Reconstitution can be achieved via a number of different strategies. The protocols for preparation of proteoliposomes use detergent removal via dialysis or via nonpolar polystyrene beads, or a mixture of the two methods. In this chapter, the protocols for nanodiscs apply polystyrene beads only. Proteoliposome preparation methods allow for substantial control of the lipid-to-protein ratio, from minimal amounts of phospholipid to high concentrations, type of phospholipid, and mixtures of phospholipids. In addition, dialysis affords a fairly large degree of control and variation of parameters such as rate of reconstitution, temperature, buffer conditions, and proteoliposome size. For the nanodisc approach, which is highly advantageous for ensuring equal access to both membrane sides of the protein as well as fast reconstitution of only a single membrane protein into a well-defined bilayer environment in each nanodisc, the protocols outline how a number of these parameters are more restricted in comparison to the proteoliposome protocols.

UI MeSH Term Description Entries
D008051 Lipid Bilayers Layers of lipid molecules which are two molecules thick. Bilayer systems are frequently studied as models of biological membranes. Bilayers, Lipid,Bilayer, Lipid,Lipid Bilayer
D008565 Membrane Proteins Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors. Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D010316 Particle Size Relating to the size of solids. Particle Sizes,Size, Particle,Sizes, Particle
D010743 Phospholipids Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system. Phosphatides,Phospholipid
D011510 Proteolipids Protein-lipid combinations abundant in brain tissue, but also present in a wide variety of animal and plant tissues. In contrast to lipoproteins, they are insoluble in water, but soluble in a chloroform-methanol mixture. The protein moiety has a high content of hydrophobic amino acids. The associated lipids consist of a mixture of GLYCEROPHOSPHATES; CEREBROSIDES; and SULFOGLYCOSPHINGOLIPIDS; while lipoproteins contain PHOSPHOLIPIDS; CHOLESTEROL; and TRIGLYCERIDES.
D003902 Detergents Purifying or cleansing agents, usually salts of long-chain aliphatic bases or acids, that exert cleansing (oil-dissolving) and antimicrobial effects through a surface action that depends on possessing both hydrophilic and hydrophobic properties. Cleansing Agents,Detergent Pods,Laundry Detergent Pods,Laundry Pods,Syndet,Synthetic Detergent,Agent, Cleansing,Agents, Cleansing,Cleansing Agent,Detergent,Detergent Pod,Detergent Pod, Laundry,Detergent Pods, Laundry,Detergent, Synthetic,Detergents, Synthetic,Laundry Detergent Pod,Laundry Pod,Pod, Detergent,Pod, Laundry,Pod, Laundry Detergent,Pods, Detergent,Pods, Laundry,Pods, Laundry Detergent,Synthetic Detergents
D003956 Dialysis A process of selective diffusion through a membrane. It is usually used to separate low-molecular-weight solutes which diffuse through the membrane from the colloidal and high-molecular-weight solutes which do not. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Dialyses
D012995 Solubility The ability of a substance to be dissolved, i.e. to form a solution with another substance. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Solubilities
D049329 Nanostructures Materials which have structured components with at least one dimension in the range of 1 to 100 nanometers. These include NANOCOMPOSITES; NANOPARTICLES; NANOTUBES; and NANOWIRES. Nanomaterials,Nanostructured Materials,Material, Nanostructured,Materials, Nanostructured,Nanomaterial,Nanostructure,Nanostructured Material

Related Publications

Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
June 2016, Nature structural & molecular biology,
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
April 2018, Journal of visualized experiments : JoVE,
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
January 2020, Methods in molecular biology (Clifton, N.J.),
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
January 1985, The Journal of membrane biology,
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
January 2017, Methods in molecular biology (Clifton, N.J.),
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
July 2002, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas,
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
January 2017, Methods in molecular biology (Clifton, N.J.),
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
January 2015, Science China. Life sciences,
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
June 2021, Biochemical Society transactions,
Kerry M Strickland, and Kasahun Neselu, and Arshay J Grant, and Carolann L Espy, and Nael A McCarty, and Ingeborg Schmidt-Krey
November 1998, Biochemistry,
Copied contents to your clipboard!