Dodecyl-β-melibioside Detergent Micelles as a Medium for Membrane Proteins

Hutchison, J.M.S.; Lu, Zhenwei; Li, Geoffrey C.; Travis, Benjamin R.; Mittal, Ritesh; Deatherage, Catherine L.; Sanders, Charles R.

doi:10.1021/acs.biochem.7b00810

Cited by 15 publications

(14 citation statements)

References 27 publications

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“…The determined size of DDM micelles corresponds well to literature [ 22 – 24 ]. However, the mean diameter of DDM micelles (7.53 nm) decreased in the presence of LIL3 (6.5 nm).…”

Section: Discussionsupporting

confidence: 86%

Characterization of chlorophyll binding to LIL3

Mork-Jansson

Eichacker

2018

PLoS ONE

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The light harvesting like protein 3 (LIL 3) from higher plants, has been linked to functions in chlorophyll and tocopherol biosynthesis, photo-protection and chlorophyll transfer. However, the binding of chlorophyll to LIL3 is unclear. We present a reconstitution protocol for chlorophyll binding to LIL3 in DDM micelles. It is shown in the absence of lipids and carotenoids that reconstitution of chlorophyll binding to in vitro expressed LIL3 requires pre-incubation of reaction partners at room temperature. We show chlorophyll a but not chlorophyll b binding to LIL3 at a molar ratio of 1:1. Neither dynamic light scattering nor native PAGE, enabled a discrimination between binding of chlorophyll a and/or b to LIL3.

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“…The determined size of DDM micelles corresponds well to literature [ 22 – 24 ]. However, the mean diameter of DDM micelles (7.53 nm) decreased in the presence of LIL3 (6.5 nm).…”

Section: Discussionsupporting

confidence: 86%

Characterization of chlorophyll binding to LIL3

Mork-Jansson

Eichacker

2018

PLoS ONE

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“…DDMB is a melibiose-derived novel detergent containing a melibiosyl moiety and a 12-carbon chain 48 (Fig. 1b ).…”

Section: Resultsmentioning

confidence: 99%

X-ray crystallography reveals molecular recognition mechanism for sugar binding in a melibiose transporter MelB

Guan

Hariharan

2021

Commun Biol

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Major facilitator superfamily_2 transporters are widely found from bacteria to mammals. The melibiose transporter MelB, which catalyzes melibiose symport with either Na+, Li+, or H+, is a prototype of the Na+-coupled MFS transporters, but its sugar recognition mechanism has been a long-unsolved puzzle. Two high-resolution X-ray crystal structures of a Salmonella typhimurium MelB mutant with a bound ligand, either nitrophenyl-α-d-galactoside or dodecyl-β-d-melibioside, were refined to a resolution of 3.05 or 3.15 Å, respectively. In the substrate-binding site, the interaction of both galactosyl moieties on the two ligands with MelBSt are virturally same, so the sugar specificity determinant pocket can be recognized, and hence the molecular recognition mechanism for sugar binding in MelB has been deciphered. The conserved cation-binding pocket is also proposed, which directly connects to the sugar specificity pocket. These key structural findings have laid a solid foundation for our understanding of the cooperative binding and symport mechanisms in Na+-coupled MFS transporters, including eukaryotic transporters such as MFSD2A.

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“…Among the cross-section of 35 detergents tested we found a number that could solubilize either Chol or SM. However, only the recently introduced 16 ndodecyl-β-melibioside (DDMB) was successful at solubilizing mixtures of various forms of phosphatidylcholine with both Chol and egg SM (eSM). eSM is predominantly (∼86%) composed of the saturated C16-amidated form (https:// avantilipids.com/product/860061) and was used instead of synthetic SM, because eSM is much less expensive.…”

Section: ■ Resultsmentioning

confidence: 99%

Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins

et al. 2020

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How the distinctive lipid composition of mammalian plasma membranes impacts membrane protein structure is largely unexplored, partly because of the dearth of isotropic model membrane systems that contain abundant sphingolipids and cholesterol. This gap is addressed by showing that sphingomyelin and cholesterol-rich (SCOR) lipid mixtures with phosphatidylcholine can be cosolubilized by n-dodecyl-β-melibioside to form bicelles. Small-angle X-ray and neutron scattering, as well as cryo-electron microscopy, demonstrate that these assemblies are stable over a wide range of conditions and exhibit the bilayered-disc morphology of ideal bicelles even at low lipid-to-detergent mole ratios. SCOR bicelles are shown to be compatible with a wide array of experimental techniques, as applied to the transmembrane human amyloid precursor C99 protein in this medium. These studies reveal an equilibrium between low-order oligomer structures that differ significantly from previous experimental structures of C99, providing an example of how ordered membranes alter membrane protein structure.

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Dodecyl-β-melibioside Detergent Micelles as a Medium for Membrane Proteins

Cited by 15 publications

References 27 publications

Characterization of chlorophyll binding to LIL3

Characterization of chlorophyll binding to LIL3

X-ray crystallography reveals molecular recognition mechanism for sugar binding in a melibiose transporter MelB

Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins

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