ArticleBiochimica et biophysica acta. Biomembranes2021
Detergent-free purification and reconstitution of functional human serotonin transporter (SERT) using diisobutylene maleic acid (DIBMA) copolymer.
Article in Biochimica et biophysica acta. Biomembranes, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 23 citations in OpenAlex.
- The Shape of Things to Come: α-Helical Membrane Protein Folding on the Ribosome.Chemical reviews · 2026Review
- Detection of an Antagonist Bound to the Neurokinin a Receptor in Styrene-Maleic Acid Lipid Particles byChembiochem : a European journal of chemical biology · 2026Article
- Cloning, Expression and Functional Characterization ofInternational journal of molecular sciences · 2025Article
- A proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs.Nature methods · 2025Article
- A proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs.bioRxiv : the preprint server for biology · 2024Article
- Identification of the potassium-binding site in serotonin transporter.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Over-Production of the Human SLC7A10 inInternational journal of molecular sciences · 2023Article
- Conformations of Human Immunodeficiency Virus Envelope Glycoproteins in Detergents and Styrene-Maleic Acid Lipid Particles.Journal of virology · 2023Article
- Lipid packing is disrupted in copolymeric nanodiscs compared with intact membranes.Biophysical journal · 2023Article
- Characterization of a soluble library of themicroLife · 2023Article
- The function of BK channels extracted and purified within SMALPs.The Biochemical journal · 2022Article
- Detergent-Free Isolation of Membrane Proteins and Strategies to Study Them in a Near-Native Membrane Environment.Biomolecules · 2022Review
- Detergent-Free Membrane Protein Purification Using SMA Polymer.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Detergent Alternatives: Membrane Protein Purification Using Synthetic Nanodisc Polymers.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Membrane protein extraction and purification using partially-esterified SMA polymers.Biochimica et biophysica acta. Biomembranes · 2021Article
- Article
- Biological insights from SMA-extracted proteins.Biochemical Society transactions · 2021Review
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3 authors at 1 institution in 1 country.
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Abstract
Structure and function analysis of human membrane proteins in lipid bilayer environments is acutely lacking despite the fundame1ntal cellular importance of these proteins and their dominance of drug targets. An underlying reason is that detailed study usually requires a potentially destabilising detergent purification of the proteins from their host membranes prior to subsequent reconstitution in a membrane mimic; a situation that is exacerbated for human membrane proteins due to the inherent difficulties in overexpressing suitable quantities of the proteins. We advance the promising styrene maleic acid polymer (SMA) extraction approach to introduce a detergent-free method of obtaining stable, functional human membrane transporters in bilayer nanodiscs directly from yeast cells. We purify the human serotonin transporter (hSERT) following overexpression in Pichia pastoris using diisobutylene maleic acid (DIBMA) as a superior method to traditional detergents or the more established styrene maleic acid polymer. hSERT plays a pivotal role in neurotransmitter regulation being responsible for the transport of the neurotransmitter 5-hydroxytryptamine (5-HT or serotonin). It is representative of the neurotransmitter sodium symporter (NSS) family, whose importance is underscored by the numerous diseases attributed to their malfunction. We gain insight into hSERT activity through an in vitro transport assay and find that DIBMA extraction improves the thermostability and activity of hSERT over the conventional detergent method.
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