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#protein extraction

3 public questions tagged with this topic.

Lipid Link Protein

What happens if a non-ionic detergent like Triton X-100 is used for extraction?

Successful extraction of functional membrane proteins relies on preserving native fold during bilayer dissolution. Non-ionic detergents like Triton X-100, n-octyl beta-D-glucoside and n-dodecyl beta-D-maltoside possess uncharged polar heads composed of polyoxyethylene chains or sugar residues. Their aliphatic tails insert between phospholipid acyl chains disrupting lipid-lipid and lipid-protein contacts while headgroups remain non-interacting with polypeptide backbone. Mixed micelles of approximately three to five nanometers surround hydrophobic transmembrane regions, extramembranous loops rem

Ref: Alberts et al., Molecular Biology of the Cell, 6th ed., Chapter 10, Non-Ionic vs Ionic Detergent Extraction.

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Lipid Link Protein

What is the role of detergents in membrane protein extraction?

Membrane protein purification requires disruption of lipid bilayer while preserving protein-protein and protein-cofactor interactions necessary for activity. Biological membranes are stabilized by hydrophobic interactions among fatty acyl chains and among transmembrane helices. Detergents are amphipathic molecules with hydrophilic head and hydrophobic tail that partition into bilayer, compete for hydrophobic surfaces, and at concentrations above critical micelle concentration form mixed micelles of lipid, detergent and protein. Appropriate non-ionic detergents like Triton X-100 and dodecyl mal

Ref: Privé, Methods 2007, Detergents for membrane protein purification; Lodish, Chapter 10.

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Membrane Transportation

What happens if a non-ionic detergent is used for membrane protein extraction?

Membrane protein extraction and purification require detergents amphipathic molecules that solubilize lipid bilayer while ideally preserving native conformation, oligomeric state and enzymatic activity for functional studies. Non-ionic detergents such as Triton X-100 octylphenoxy polyoxyethylene, n-dodecyl-beta-D-maltoside DDM, digitonin steroidal glycoside and n-octyl-beta-D-glucoside possess uncharged hydrophilic head groups composed of polyoxyethylene or sugar moieties and hydrophobic alkyl tail. They insert into bilayer at low critical micellar concentration, fragment it into mixed lipid-d

Ref: Le Maire et al., Biochimica et Biophysica Acta 2000: Non-Ionic Detergents Preserve Membrane Protein Folding.

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