Biophysical Characterization of Membrane Proteins

Niebling, Stephan; Burastero, Osvaldo; García-Alai, María

doi:10.1007/978-1-0716-3147-8_12

Cited by 4 publications

(2 citation statements)

References 32 publications

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“…MST is a relatively simple, robust, and rapid technique that has several advantages over other biophysical techniques commonly used to measure biomolecular interactions such as surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), dynamic light scattering (DLS), fluorescence spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and mass spectrometry (MS) [6,9,[11][12][13][14][15][16][17][18][19][20]. MST has no limitations on the size of the target biomolecule or on the affinity of the interaction to be measured (pM to mM) or on the composition of the buffer and other sample components.…”

Section: Advantages Of Mstmentioning

confidence: 99%

Microscale Thermophoresis Analysis of Membrane Proteins

Patching

2024

Preprint

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Microscale thermophoresis (MST) is an analytical technique for measuring biomolecular interactions. It is based on the physical phenomenon that particles move within temperature gradients, which is affected by their size, charge, hydration shell and conformation. The MST sample must contain a fluorescent target molecule used to observe the movement of particles, and this can be titrated with an unlabelled binding partner for quantifying the interaction. MST is highly sensitive, using relatively small amounts of sample, and it has no limitations on the size of the target biomolecule, on the affinity of the interaction or on the composition of the buffer and other sample components. This makes MST ideally suited to characterising interactions with membrane proteins, which can be studied in cell lysates, native membranes, solubilised in detergents or reconstituted in lipids. The intrinsic aromatic residues of membrane proteins have been used as the fluorophore for MST (label-free MST) or membrane proteins have been labelled with a range of fluorescent dyes or conjugated with fluorescent proteins (labelled MST). The different types of membrane proteins that have had biomolecular interactions characterised by MST include the SARS-CoV-2 spike protein, GPCRs and other receptors, sensor kinases, ion channels, aquaporins, and transport proteins.

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Section: Advantages Of Mstmentioning

confidence: 99%

Microscale Thermophoresis Analysis of Membrane Proteins

Patching

2024

Preprint

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“…Detergents are one of the most widely used reagents in molecular biology research [1][2][3]. Their use includes but is not limited to cell lysis, electrophoresis, protein refolding, and the suppression of macromolecular surface adsorption or aggregation, i.e., a manipulation of macromolecular solutions [4].…”

Section: Introductionmentioning

confidence: 99%

Sodium Dodecyl Sulfate Analogs as a Potential Molecular Biology Reagent

Arakawa,

Niikura,

Kita

et al. 2024

CIMB

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In this study, we review the properties of three anionic detergents, sodium dodecyl sulfate (SDS), Sarkosyl, and sodium lauroylglutamate (SLG), as they play a critical role in molecular biology research. SDS is widely used in electrophoresis and cell lysis for proteomics. Sarkosyl and, more frequently, SDS are used for the characterization of neuropathological protein fibrils and the solubilization of proteins. Many amyloid fibrils are resistant to SDS or Sarkosyl to different degrees and, thus, can be readily isolated from detergent-sensitive proteins. SLG is milder than the above two detergents and has been used in the solubilization and refolding of proteins isolated from inclusion bodies. Here, we show that both Sarkosyl and SLG have been used for protein refolding, that the effects of SLG on the native protein structure are weaker for SLG, and that SLG readily dissociates from the native proteins. We propose that SLG may be effective in cell lysis for functional proteomics due to no or weaker binding of SLG to the native proteins.

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