Lipid-Detergent Phase Transitions During Detergent-Mediated Liposome Solubilization

Niroomand, Hanieh; Venkatesan, Guru A.; Sarles, Stephen A.; Mukherjee, Dibyendu; Khomami, Bamin

doi:10.1007/s00232-016-9894-1

Cited by 22 publications

(21 citation statements)

References 73 publications

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“…Having established ensemble FRET as an optical sensor of changes in dye separation distance as a function of SDS, DLS was next used to probe the hydrodynamic radii of LUVs in response to SDS in solution. The high sensitivity of DLS to the diffusion of nanomolar concentrations of vesicles makes it an attractive technique for accessing structural characteristics along the solubilization pathway [25,26] . Here, the autocorrelation function obtained from the intensity of light scattered by the LUVs yields their translational diffusion coefficient which can be used to extrapolate their hydrodynamic radius (rH) via the Stokes-Einstein equation.…”

Section: Resultsmentioning

confidence: 99%

The Mechanism of Vesicle Solubilization by the Detergent Sodium Dodecyl Sulfate

Juan-Colás

Dresser

Morris

et al. 2020

Preprint

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Membrane solubilization by sodium dodecyl sulfate (SDS) is indispensable for many established biotechnological applications, including viral inactivation and protein extraction. Although the ensemble thermodynamics have been thoroughly explored, the underlying molecular dynamics have remained inaccessible, owing to major limitations of traditional measurement tools. Here, we integrate multiple advanced biophysical approaches to gain multi-angle insight into the time-dependence and fundamental kinetic steps associated with the solubilization of single sub-micron sized vesicles in response to SDS. We find that the accumulation of SDS molecules on in-tact vesicles triggers biphasic solubilization kinetics comprising an initial vesicle expansion event followed by rapid lipid loss and micellization. Our findings support a general mechanism of detergent-induced membrane solubilization and we expect the framework of correlative biophysical technologies presented here will form a general platform for elucidating the complex kinetics of membrane perturbation induced by a wide variety of surfactants and disrupting agents.

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Section: Resultsmentioning

confidence: 99%

The Mechanism of Vesicle Solubilization by the Detergent Sodium Dodecyl Sulfate

Juan-Colás

Dresser

Morris

et al. 2020

Preprint

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“…Reconstitution of integral membrane proteins typically involves insertion into either mixed micelles or preformed vesicles. The former comprises mixing of detergent-lipid and detergent-protein-lipid micelles, with spontaneous bilayer formation and protein insertion occurring during the detergent removal phase [38,46,47]. However, this route may result in an unpredictable size distribution of proteoliposomes and the presence of multilamellar structures.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of Hybrid Polymersome Vesicles Containing the Efflux Pumps NaAtm1 or P-Glycoprotein

et al. 2020

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Investigative systems for purified membrane transporters are almost exclusively reliant on the use of phospholipid vesicles or liposomes. Liposomes provide an environment to support protein function; however, they also have numerous drawbacks and should not be considered as a “one-size fits all” system. The use of artificial vesicles comprising block co-polymers (polymersomes) offers considerable advantages in terms of structural stability; provision of sufficient lateral pressure; and low passive permeability, which is a particular issue for transport assays using hydrophobic compounds. The present investigation demonstrates strategies to reconstitute ATP binding cassette (ABC) transporters into hybrid vesicles combining phospholipids and the block co-polymer poly (butadiene)-poly (ethylene oxide). Two efflux pumps were chosen; namely the Novosphingobium aromaticivorans Atm1 protein and human P-glycoprotein (Pgp). Polymersomes were generated with one of two lipid partners, either purified palmitoyl-oleoyl-phosphatidylcholine, or a mixture of crude E. coli lipid extract and cholesterol. Hybrid polymersomes were characterised for size, structural homogeneity, stability to detergents, and permeability. Two transporters, NaAtm1 and P-gp, were successfully reconstituted into pre-formed and surfactant-destabilised hybrid polymersomes using a detergent adsorption strategy. Reconstitution of both proteins was confirmed by density gradient centrifugation and the hybrid polymersomes supported substrate dependent ATPase activity of both transporters. The hybrid polymersomes also displayed low passive permeability to a fluorescent probe (calcein acetomethoxyl-ester (C-AM)) and offer the potential for quantitative measurements of transport activity for hydrophobic compounds.

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“…The highly irregular structural arrangements that arises in this system at room temperature indicates its unsuitability for successful protein insertion. Furthermore, this study unveiled the possibility of 1,2-diphytanoyl- sn -glycero-3-phosphocholine (DPhPC) as a promising lipid candidate for protein insertions 27 . It should be noted that DPhPC is a branched phosphatidylcholine (PC) lipid with high thermal and structural stability 28 .…”

Section: Introductionmentioning

confidence: 90%

Tuning the photoexcitation response of cyanobacterial Photosystem I via reconstitution into Proteoliposomes

Niroomand

Mukherjee

Khomami

2017

Sci Rep

Self Cite

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The role of natural thylakoid membrane housing of Photosystem I (PSI), the transmembrane photosynthetic protein, in its robust photoactivated charge separation with near unity quantum efficiency is not fundamentally understood. To this end, incorporation of suitable protein scaffolds for PSI incorporation is of great scientific and device manufacturing interest. Areas of interest include solid state bioelectronics, and photoelectrochemical devices that require bio-abio interfaces that do not compromise the photoactivity and photostability of PSI. Therefore, the surfactant-induced membrane solubilization of a negatively charged phospholipid (DPhPG) with the motivation of creating biomimetic reconstructs of PSI reconstitution in DPhPG liposomes is studied. Specifically, a simple yet elegant method for incorporation of PSI trimeric complexes into DPhPG bilayer membranes that mimic the natural thylakoid membrane housing of PSI is introduced. The efficacy of this method is demonstrated via absorption and fluorescence spectroscopy measurements as well as direct visualization using atomic force microscopy. This study provides direct evidence that PSI confinements in synthetic lipid scaffolds can be used for tuning the photoexcitation characteristics of PSI. Hence, it paves the way for development of fundamental understanding of microenvironment alterations on photochemical response of light activated membrane proteins.

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Lipid-Detergent Phase Transitions During Detergent-Mediated Liposome Solubilization

Cited by 22 publications

References 73 publications

The Mechanism of Vesicle Solubilization by the Detergent Sodium Dodecyl Sulfate

The Mechanism of Vesicle Solubilization by the Detergent Sodium Dodecyl Sulfate

Characterisation of Hybrid Polymersome Vesicles Containing the Efflux Pumps NaAtm1 or P-Glycoprotein

Tuning the photoexcitation response of cyanobacterial Photosystem I via reconstitution into Proteoliposomes

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