Studies on the mechanism of membrane fusion: kinetics of calcium ion induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents

Wilschut, Jan; Düzgüneş, Nejat; Fraley, Robert T.; Papahadjopoulos, Demetrios

doi:10.1021/bi00567a011

Cited by 493 publications

(400 citation statements)

References 52 publications

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“…DOPC vesicles with each of these fluorophores were prepared and tested for their usefulness in the assay. The Tb 3+ /DPA system, which has been used in several biological applications such as endospore detection [36], vesicle fusion [37,38], and vesicle leakage [38,39], proved to be superior to the fluo-3 and calcium green 1 systems. The latter fluorophores are sensitive to calcium ions, and there…”

Section: Evaluation Of Fluorescence Systemsmentioning

confidence: 99%

“…Second, in the absence of a chelator, Tb 3+ has been shown to change the fluidity characteristics of phosphatidylserine (PS) when encapsulated in PS vesicles [37]. Citrate was used as a chelator in the experiments in order to minimize the association of Tb 3+ with the lipids, but the use of CF avoided this complication entirely.…”

Section: Evaluation Of Fluorescence Systemsmentioning

confidence: 99%

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Mechanosensitive channel of large conductance

Kloda

Petrov

Meyer

et al. 2008

The International Journal of Biochemistry & Cell Biology

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The bacterial mechanosensitive channel of large conductance (MscL) is an ideal starting point for understanding the molecular basis of mechanosensation. However, current methods for the characterization of its mutants, patch clamp and bacterial growth analysis, are difficult and time consuming, so a higher throughput method for screening mutants is desired. We have attempted to develop a fluorescence assay for detecting MscL activity in synthetic vesicles. The assay involved the separation of two solutionsone inside and one outside the vesicles-that are separately nonfluorescent but fluorescent when mixed. It was hoped that MscL activity due to downshock of the vesicles would bring about mixing of the solutions, producing fluorescence. The development of the assay required the optimization of several variables: the method for producing a uniform vesicle population containing MscL, the fluorescence system, and the lipid and protein composition of the vesicles. However, no MscL activity was ever detected even after optimization, so the assay was not fully developed. The probable cause of the failure was the inability of current techniques to produce a sufficiently uniform vesicle population.

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Section: Evaluation Of Fluorescence Systemsmentioning

confidence: 99%

Section: Evaluation Of Fluorescence Systemsmentioning

confidence: 99%

Mechanosensitive channel of large conductance

Kloda

Petrov

Meyer

et al. 2008

The International Journal of Biochemistry & Cell Biology

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“…The prerequisite of membrane contact to the destabilization of anionic membranes in the presence of divalent cations was demonstrated by a number of workers (Liao & Pestegard, 1980b;Liao & Pestegard, 1980a;Wilschut et al, 1980;Nir et al, 1983a). Later, Ellens et al (Ellens et al, 1984) showed that membrane contact was also necessary for destabilization of PE-containing liposomes.…”

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confidence: 99%

“…In the Tb/DPA assay, separate liposome populations are prepared containing either Tb 3 + or DPA2- (Wilschut & Papahadjopoulos, 1979;Wilschut et al, 1980). Both species exhibit high solubilities in aqueous buffers due to their charges.…”

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confidence: 99%

Photoinduced Fusion of Liposomes

Bennett¹,

O’Brien²

1994

J. Am. Chem. Soc.

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This manuscript ,has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissenation copies are in typewriter face, while others may be from any type of computer printer.The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction.In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note wiD indicate the deletion.Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continning from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book.

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“…[1][2][3][4][5][6][7][8] Liposomes can be considered as a model of protocells in studies on the origin of life as they are readily prepared in the aqueous environment: the lipid bilayer system produced around clay particles might have evolved into a kind of protocell in primeval sea that separated from the clay. 9) The interaction between lecithin liposomes and clay has been investigated by one of the present authors 10) who found that liposomes were adsorbed to bentonite, a kind of montmorillonite, and the clay particles aggregated to sediment as a result of this adsorption.…”

mentioning

confidence: 99%