scholarly journals Interaction of cholesterol-crystallization-promoting proteins with vesicles

1995 ◽  
Vol 305 (1) ◽  
pp. 93-96 ◽  
Author(s):  
M A C de Bruijn ◽  
B G Goldhoorn ◽  
A I M Zijlstra ◽  
G N J Tytgat ◽  
A K Groen
Keyword(s):  
Energy Transfer ◽  
Concanavalin A ◽  
Binding Proteins ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Phospholipid Vesicles ◽  
Human Bile ◽  
Induced Crystallization

In this study, the interaction of mucin and concanavalin A-binding proteins isolated from human bile with cholesterol/phospholipid vesicles was investigated. Using resonance energy transfer assays originally developed by Struck, Hoekstra and Pagano [(1981) Biochemistry 20, 4093-4099], no significant protein-induced fusion or aggregation of vesicles was demonstrated. Instead of fusion, these proteins induced destabilization of cholesterol/phospholipid vesicles, as monitored by release of entrapped carboxyfluorescein. A good correlation (rho = 0.81) was obtained between the extent of leakage and the nucleation-promoting activity of the concanavalin A-binding proteins. We conclude that aggregation or fusion of cholesterol/phospholipid vesicles is not an obligatory step in cholesterol crystallization. Biliary protein-induced crystallization seems to be preceded by vesicle disruption.

scholarly journals Dibucaine interaction with phospholipid vesicles. A resonance energy-transfer study

1990 ◽  
Vol 189 (2) ◽  
pp. 387-393 ◽  
Author(s):  
Ana COUTINHO ◽  
Julia COSTA ◽  
Joaquim L. FARIA ◽  
Mario N. BERBERAN-SANTOS ◽  
Manuel J. E. PRIETO
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Phospholipid Vesicles ◽  
Transfer Study

A biosensing platform for sensitive detection of concanavalin A based on fluorescence resonance energy transfer from CdTe quantum dots to graphene oxide

2015 ◽  
Vol 39 (8) ◽  
pp. 6092-6098 ◽  
Author(s):  
Yan Li ◽  
Fanping Shi ◽  
Nan Cai ◽  
Xingguang Su
Keyword(s):  
Quantum Dots ◽  
Graphene Oxide ◽  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Concanavalin A ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Cdte Quantum Dots ◽  
Sensitive Detection ◽  
Sandwich Method

The sandwich method can detect different lectins simply by exchanging the carbohydrates functionalized on the quantum dots and graphene oxide.

scholarly journals Design and Application of Highly Responsive Fluorescence Resonance Energy Transfer Biosensors for Detection of Sugar in Living Saccharomyces cerevisiae Cells

2007 ◽  
Vol 73 (22) ◽  
pp. 7408-7414 ◽  
Author(s):  
Jae-Seok Ha ◽  
Jae Jun Song ◽  
Young-Mi Lee ◽  
Su-Jin Kim ◽  
Jung-Hoon Sohn ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Signal Intensity ◽  
Binding Proteins ◽  
Dynamic Range ◽  
Binding Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy

ABSTRACT A protein sensor with a highly responsive fluorescence resonance energy transfer (FRET) signal for sensing sugars in living Saccharomyces cerevisiae cells was developed by combinatorial engineering of the domain linker and the binding protein moiety. Although FRET sensors based on microbial binding proteins have previously been created for visualizing various sugars in vivo, such sensors are limited due to a weak signal intensity and a narrow dynamic range. In the present study, the length and composition of the linker moiety of a FRET-based sensor consisting of CFP-linker1-maltose-binding protein-linker2-YFP were redesigned, which resulted in a 10-fold-higher signal intensity. Molecular modeling of the composite linker moieties, including the connecting peptide and terminal regions of the flanking proteins, suggested that an ordered helical structure was preferable for tighter coupling of the conformational change of the binding proteins to the FRET response. When the binding site residue Trp62 of the maltose-binding protein was diversified by saturation mutagenesis, the Leu mutant exhibited an increased binding constant (82 μM) accompanied by further improvement in the signal intensity. Finally, the maltose sensor with optimized linkers was redesigned to create a sugar sensor with a new specificity and a wide dynamic range. When the optimized maltose sensors were employed as in vivo sensors, highly responsive FRET images were generated from real-time analysis of maltose uptake of Saccharomyces cerevisiae (baker's yeast).

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