Use of energy transfer to assay the asseciation of proteins with lipid membranes

Vaz, Winchil L.C.; Kaufmann, Konrad; Nicksch, Alfar

doi:10.1016/0003-2697(77)90047-1

Cited by 30 publications

(10 citation statements)

References 9 publications

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“…Note the systematic decrease in the enzyme's fluorescence emission intensity ( λ em = 343 nm) with concomitant increase in the intensity of the dansyl peak (at 526 nm). The overall spectral changes conform to a single isosbestic point at 397 nm, consistent with the fluorescence resonance energy transfer (FRET) from the excited state of the enzyme's tryptophan/tyrosine residues to the liposome‐resident dansyl probe, and its origin must lie in the formation of the enzyme‐liposome complex [12]. In addition, the dissociation constants calculated from the liposome concentration‐dependent changes in the fluorescence intensities at 343 nm (due to quenching of the enzyme's fluorescence) and at 526 nm (due to enhancement of the dansyl fluorescence) were found to be remarkably similar, e.g., 0.73 μM and 0.87 μM, respectively.…”

Section: Resultssupporting

confidence: 65%

“…The binding of MMP‐7 to differently charged liposomes was determined by measuring the decrease in the protein's intrinsic fluorescence ( λ ex = 283 nm, λ em = 343 nm) and the concurrent increase in the fluorescence of the lipid probe as a function of increasing lipid concentration. 1,2‐Dioleoyl‐ sn ‐glycero‐3‐phosphoethanolamine‐ N (5‐dimethylamino‐1‐naphthalenesulfonyl) lipid (dansyl‐PE) ( λ em = 526 nm, Avanti ® Polar Lipids, Alabaster, AL) was utilized as the liposome incorporated fluorescence reporter group [12]. All fluorescence measurements were performed using a LS‐50B Perkin–Elmer ® spectroflourimeter.…”

Section: Methodsmentioning

confidence: 99%

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Intrinsic selectivity in binding of matrix metalloproteinase‐7 to differently charged lipid membranes

et al. 2007

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We provide evidence that matrix metalloproteinase-7 (MMP-7) interacts with anionic, cationic and neutral lipid membranes, although it interacts strongest with anionic membranes. While the catalytic activity of the enzyme remains unaffected upon binding to neutral and negatively charged membranes, it is drastically impaired upon binding to the positively charged membranes. The structural data reveal that the origin of these features lies in the ''bipolar'' distribution of the electrostatic surface potentials on the crystallographic structure of MMP-7.

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Section: Resultssupporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Intrinsic selectivity in binding of matrix metalloproteinase‐7 to differently charged lipid membranes

et al. 2007

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“…Dansylated PE molecules, containing a dansyl fluorophore covalently linked to the primary amine of the ethanolamine of PE, were incorporated in small (usually 5 mol %) proportions into lipid vesicles. Endonexin-membrane binding was then detected by the distance-dependent energy transfer between excited endonexin aromatic residues and the dansyl fluorophores localized to the vesicles (Bazzi & Nelsestuen, 1987;Vaz et al, 1977). Fluorescence measurements were performed on a SPEX Fluorolog 2 fluorometer with excitation at 280 nm and emission monitored at 510 nm.…”

Section: Methodsmentioning

confidence: 99%

Ca2+-Dependent Binding of Endonexin (Annexin IV) to Membranes: Analysis of the Effects of Membrane Lipid Composition and Development of a Predictive Model for the Binding Interaction

Junker¹,

Creutz²

1994

Biochemistry

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Endonexin (annexin IV) is a member of the annexin family of homologous proteins that bind membranes and aggregate vesicles in a calcium-dependent fashion. This study examines the lipid modulation and mechanism of the binding of endonexin to membranes using a fluorescence energy transfer assay to measure bovine endonexin binding to well-defined large unilamellar vesicles. The calcium sensitivity for endonexin-membrane binding is observed to be highly dependent on the types of membrane lipids present. As with most annexins, negatively charged lipids best promote endonexin binding to phosphatidylcholine (PC) containing membranes. However, a comparison of 11 different types of lipids reveals that other factors such as the type of ion contributing the charge and head-group size are also important. The concentrations of calcium required for half-maximal binding of endonexin to PC vesicles containing 30% phosphatidylserine (PS) or 30% phosphatidylinositol (PI), both lipids with net charge-1, are 48 +/- 6 and 114 +/- 19 microM, respectively, while half-maximal binding to 30% phosphatidylinositol bisphosphate (PIP2), with a greater net charge of -3 to -5, occurs at 65 microM calcium, similar to the calcium requirement for binding to PS. The apparent affinities of endonexin for seven different types of lipids parallel those reported for annexin V [Andree, H. A. M., Reutelingsperger, C. P. M., Hauptmann, R., Hemker, H. C., Hermans, W. T., & Willems, G. M. (1990) J. Biol. Chem. 265, 4923-4928], except for a greater preference of endonexin for membranes containing phosphatidic acid. Mixing PS and phosphatidylethanolamine (PE) or PS and PI in the same PC vesicle synergistically enhances endonexin-membrane binding, indicating that even lipids with no net charge such as PE may dramatically affect endonexin binding to mixed-lipid membranes. The maximum amount of endonexin able to bind to PS/PC vesicles at 1 mM calcium increases with mole % PS. A simple and general model that treats protein-membrane binding as a two-step process, with adsorption to a membrane surface followed by interaction with specific lipid molecules [Lentz, B. R., & Hermans, J. (1989) Biochemistry 28, 7459-7461], is extended to include the coupled binding of calcium with binding of specific lipid molecules. This extended model accurately predicts trends observed when protein and calcium titrations of endonexin binding to PS/PC vesicles are performed under a wide variety of conditions and suggests that 3-5 calcium ions and 9-18 PS molecules participate in each endonexin-membrane complex.(ABSTRACT TRUNCATED AT 400 WORDS)

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“…The steady-state fluorimetric technique was extensively for monitoring Ca 2+promoted vesicle-vesicle, chromaffin granule fusion (Struck et al 1981;Morris et al 1982 a,*;Morris & Bradley, 1984) and poly(ethylene glycol)-induced fusion of phospholipid vesicles (Morgan et al 1983). FRET measurements were applied for the determination of the size of the protein-lipid complex in membrane (Vaz et al 1977) of the size of the protein induced perturbation of the lipid phase (Rehorek et al 1985). FRET was used to characterize how deep the haem moiety of cytochrome b b is localized from the surface of the membrane (Kleinfeld & Lukacovich, 1985) what is the aggregation state of bacteriorhodopsin (Hasselbacher et al 1984), sarcoplasmic reticulum ATP-ase (Vanderkooi et al 19JT, Papp et al 1987) and membrane-bound complexes of complement proteins (Cheng et al 1985).…”

Section: Biological Significance Of Fret and Fcet Measurementsmentioning

confidence: 99%

Luminescence spectroscopic approaches in studying cell surface dynamics

Matkó

Szöllősi

Trón

et al. 1988

Quart. Rev. Biophys.

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The major elements of membranes, such as proteins, lipids and polysaccharides, are in dynamic interaction with each other (Albertset al.1983). Protein diffusion in the lipid matrix of the membrane, the lipid diffusion and dynamic domain formation below and above their transition temperature from gel to fluid state, have many functional implications. This type of behaviour of membranes is often summarized in one frequently used word membrane fluidity (coined by Shinitzky & Henkart, 1979). The dynamic behaviour of the cell membrane includes rotational, translational and segmental movements of membrane elements (or their domain-like associations) in the plane of, and perpendicular to the membrane. The ever changing proximity relationships form a dynamic pattern of lipids, proteins and saccharide moieties and are usually described as ‘cell-surface dynamics’ (Damjanovichet al.1981). The knowledge about the above defined behaviour originates from experiments performed mostly on cytoplasmic membranes of eukaryotic cells. Nevertheless numerous data are available also on the mitochondrial and nuclear membranes, as well as endo (sarco-)plasmic reticulum (Martonosi, 1982; Slater, 1981; Siekevitz, 1981).

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Use of energy transfer to assay the asseciation of proteins with lipid membranes

Cited by 30 publications

References 9 publications

Intrinsic selectivity in binding of matrix metalloproteinase‐7 to differently charged lipid membranes

Intrinsic selectivity in binding of matrix metalloproteinase‐7 to differently charged lipid membranes

Ca2+-Dependent Binding of Endonexin (Annexin IV) to Membranes: Analysis of the Effects of Membrane Lipid Composition and Development of a Predictive Model for the Binding Interaction

Luminescence spectroscopic approaches in studying cell surface dynamics

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