Surface plasmon resonance in protein–membrane interactions

Beseničar, Mojca Podlesnik; Maček, Peter; Lakey, Jeremy H.; Anderluh, Gregor

doi:10.1016/j.chemphyslip.2006.02.010

Cited by 206 publications

(150 citation statements)

References 84 publications

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“…This technique is used systematically to study various biomolecular interactions with both lipids and proteins (Besenicar et al, 2006). In the Lti30 and phospholipid recognition studies, the phospholipid vesicles were immobilized on sensor chip surfaces, and the dehydrins served as the soluble analytes.…”

Section: Dehydrin-phospholipid Interaction Studies: Surface Plasmon Rmentioning

confidence: 99%

Tunable Membrane Binding of the Intrinsically Disordered Dehydrin Lti30, a Cold-Induced Plant Stress Protein

et al. 2011

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Dehydrins are intrinsically disordered plant proteins whose expression is upregulated under conditions of desiccation and cold stress. Their molecular function in ensuring plant survival is not yet known, but several studies suggest their involvement in membrane stabilization. The dehydrins are characterized by a broad repertoire of conserved and repetitive sequences, out of which the archetypical K-segment has been implicated in membrane binding. To elucidate the molecular mechanism of these K-segments, we examined the interaction between lipid membranes and a dehydrin with a basic functional sequence composition: Lti30, comprising only K-segments. Our results show that Lti30 interacts electrostatically with vesicles of both zwitterionic (phosphatidyl choline) and negatively charged phospholipids (phosphatidyl glycerol, phosphatidyl serine, and phosphatidic acid) with a stronger binding to membranes with high negative surface potential. The membrane interaction lowers the temperature of the main lipid phase transition, consistent with Lti30's proposed role in cold tolerance. Moreover, the membrane binding promotes the assembly of lipid vesicles into large and easily distinguishable aggregates. Using these aggregates as binding markers, we identify three factors that regulate the lipid interaction of Lti30 in vitro: (1) a pH dependent His on/off switch, (2) phosphorylation by protein kinase C, and (3) reversal of membrane binding by proteolytic digest.

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Section: Dehydrin-phospholipid Interaction Studies: Surface Plasmon Rmentioning

confidence: 99%

Tunable Membrane Binding of the Intrinsically Disordered Dehydrin Lti30, a Cold-Induced Plant Stress Protein

et al. 2011

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“…We investigate transport dynamics in a model geometry of a surface plasmon resonance (SPR) sensor, which by far is the most common SPR platform used. 24 The finite distance from the inlet of the microfluidic flow cell to the sensor surface is included, as shown in Figure 1. The length scales are l = 2.3 mm, w = 0.5 mm, and h = 0.02 mm.…”

Section: ■ Theorymentioning

confidence: 99%

Transient Convection, Diffusion, and Adsorption in Surface-Based Biosensors

et al. 2012

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This paper presents a theoretical and computational investigation of convection, diffusion, and adsorption in surface-based biosensors. In particular, we study the transport dynamics in a model geometry of a surface plasmon resonance (SPR) sensor. The work, however, is equally relevant for other microfluidic surface-based biosensors, operating under flow conditions. A widely adopted approximate quasi-steady theory to capture convective and diffusive mass transport is reviewed, and an analytical solution is presented. An expression of the Damkoḧler number is derived in terms of the nondimensional adsorption coefficient (Biot number), the nondimensional flow rate (Pećlet number), and the model geometry. Transient dynamics is investigated, and we quantify the error of using the quasi-steady-state assumption for experimental data fitting in both kinetically limited and convection-diffusionlimited regimes for irreversible adsorption, in specific. The results clarify the conditions under which the quasi-steady theory is reliable or not. In extension to the well-known fact that the range of validity is altered under convection-diffusion-limited conditions, we show how also the ratio of the inlet concentration to the maximum surface capacity is critical for reliable use of the quasi-steady theory. Finally, our results provide users of surface-based biosensors with a tool for correcting experimentally obtained adsorption rate constants.

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“…SPR, in particular, has proven to be a valuable experimental approach in the studies of protein membrane binding (40)(41)(42). Liposomes are immobilized on the surface of the sensor chip, and the protein of interest is passed across them.…”

Section: Binding Of Llo To Immobilized Liposomesmentioning

confidence: 99%

Sterol and pH Interdependence in the Binding, Oligomerization, and Pore Formation of Listeriolysin O

et al. 2007

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Listeriolysin O (LLO) is the most important virulence factor of the intracellular pathogen Listeria monocytogenes. Its main task is to enable escape of bacteria from the phagosomal vacuole into the cytoplasm. LLO belongs to the cholesterol-dependent cytolysin (CDC) family but differs from other members, as it exhibits optimal activity at low pH. Its pore forming ability at higher pH values has been largely disregarded in Listeria pathogenesis. Here we show that high cholesterol concentrations in the membrane restore the low activity of LLO at high pH values. LLO binds to lipid membranes, at physiological or even slightly basic pH values, in a cholesterol-dependent fashion. Binding, insertion into lipid monolayers, and permeabilization of calcein-loaded liposomes are maximal above approximately 35 mol % cholesterol, a concentration range typically found in lipid rafts. The narrow transition region of cholesterol concentration separating low and high activity indicates that cholesterol not only allows the binding of LLO to membranes but also affects other steps in pore formation. We were able to detect some of these by surface plasmon resonance-based assays. In particular, we show that LLO recognition of cholesterol is determined by the most exposed 3 -hydroxy group of cholesterol. In addition, LLO binds and permeabilizes J774 cells and human erythrocytes in a cholesterol-dependent fashion at physiological or slightly basic pH values. The results clearly show that LLO activity at physiological pH cannot be neglected and that its action at sites distal to cell entry may have important physiological consequences for Listeria pathogenesis.

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Surface plasmon resonance in protein–membrane interactions

Cited by 206 publications

References 84 publications

Tunable Membrane Binding of the Intrinsically Disordered Dehydrin Lti30, a Cold-Induced Plant Stress Protein

Tunable Membrane Binding of the Intrinsically Disordered Dehydrin Lti30, a Cold-Induced Plant Stress Protein

Transient Convection, Diffusion, and Adsorption in Surface-Based Biosensors

Sterol and pH Interdependence in the Binding, Oligomerization, and Pore Formation of Listeriolysin O

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