Characterization of nanodisc-forming peptides for membrane protein studies

Nanodisc Reconstitution and Characterization of Amyloid-β Precursor Protein C99

“…Most of the reported nanodisc-forming synthetic polymers are ionic, and hence, they can interact with the desired membrane protein(s) to be reconstituted in nanodiscs, affecting their structure and function. Such undesired effects of the use of charged polymers have been reported for the positively charged cytochrome-P450 and the negatively charged cytochrome-b5 proteins, and the difficulty in functionally reconstituting P450-b5 complex. , A similar observation has been reported for the nanodisc-forming cationic-rich AEM28 peptide . Thus, charged polymers/peptides are not suitable to study membrane proteins (or other biomolecules) that can exhibit electrostatic interactions with the nanodisc-forming polymers/peptides.…”

“…51,52 A similar observation has been reported for the nanodisc-forming cationic-rich AEM28 peptide. 39 Thus, charged polymers/ peptides are not suitable to study membrane proteins (or other biomolecules) that can exhibit electrostatic interactions with the nanodisc-forming polymers/peptides. To overcome these limitations, we developed a nonionic pentyl-inulin polymer by functionalizing inulin.…”

“…Although detergents are cost-effective and efficient in extracting membrane proteins, they have limitations, as their short alkyl chains and headgroups are chemically distinct from those of native membrane lipids, rendering them discrete physicochemical properties that can affect the function of membrane proteins. ,− Moreover, their denaturing properties make it difficult to maintain membrane protein’s oligomeric or multimeric states that are stabilized by intermolecular interactions. − Planar lipid-bilayer-containing bicelles have been used extensively to study the structure, dynamics, and membrane topology of a variety of membrane proteins . However, bicelles also contain detergent-like short-chain lipids that could affect the stability of a reconstituted membrane protein. − Likewise, liposomes are also limited by a lack of good stability and vesicular morphology. ,, Detergent-based membrane-mimicking systems offer minimal stability , and may not be desirable for high-resolution studies of membrane protein. , On the other hand, nanodiscs, which are nanometer-sized lipid bilayered systems surrounded and stabilized by a protective belt of peptides, proteins, or polymers , have been shown to provide a stable and more native-like membrane environment for studying the structure and function of various membrane proteins. ,− …”

Novel class of peptides disintegrating biological membranes to aid in the characterization of membrane proteins

“…In our recent experiments, we found that the FFD8 peptide C-terminally modified with FLAG tag (DYKDDDDK) or N-terminally with biotin also disintegrated Jurkat cell membranes (unpublished results). It should be noted that a recent paper reports membrane-disintegrating properties of another type of peptides ( 18 ). Last, it may be speculated that FFDn-like peptides encoded in an expression vector with specific (e.g.…”