Direct observation of the growth and shrinkage of microtubules by single molecule Förster resonance energy transfer

Mondal, Prasenjit; Chowdhury, Rajdeep; Bhunia, Debmalya; Ghosh, Surajit; Bhattacharyya, Kankan

doi:10.1039/c4cp06031h

Cited by 12 publications

(26 citation statements)

References 44 publications

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“…Time dependent smFRET efficiency fluctuations of antigen-antibody complex show a broad range from 0.2 to 0.9, corresponding to distance fluctuates from 2 to 7.5 nm. 84,85 …”

Section: Applications Of Smfretmentioning

confidence: 99%

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Single-molecule fluorescence resonance energy transfer in molecular biology

et al. 2016

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Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful technique for studying the conformation dynamics and interactions of individual biomolecules. In this review, we describe the concept and principle of smFRET, illustrate general instrumentation and microscopy settings for experiments, and discuss the methods and algorithms for data analysis. Subsequently, we review applications of smFRET in protein conformational changes, ion channel open-close properties, receptor-ligand interactions, nucleic acid structure regulation, vesicle fusion, and force induced conformational dynamics. Finally, we discuss the main limitations of smFRET in molecular biology.

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“…Time dependent smFRET efficiency fluctuations of antigen-antibody complex show a broad range from 0.2 to 0.9, corresponding to distance fluctuates from 2 to 7.5 nm. 84,85 …”

Section: Applications Of Smfretmentioning

confidence: 99%

“…Figures are reproduced from Mandal et al with permission. 85 Copyright (2015) Royal Society of Chemistry.…”

Section: Figmentioning

confidence: 99%

Single-molecule fluorescence resonance energy transfer in molecular biology

et al. 2016

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“…We have used sm‐FRET to investigate the growth (polymerization) and shrinkage (depolymerization) of the dynamic microtubules . Microtubules are polar, long, tube‐like dynamic polymers of αβ‐tubulin heterodimer.…”

Section: Conformational Fluctuation Of Proteins: Fcs and Sm‐fretmentioning

confidence: 99%

“…Dynamic structural fluctuations of microtubules play a crucial role in cell division, maintaining intracellular space and traffcking in eukaryotic cells. For the sm‐FRET study, we used EGFP (attached to Mal3) as a donor and Alexa 568 bound to tubulin as an acceptor . Mal3 binds to microtubulin only when it is polymerized.…”

Section: Conformational Fluctuation Of Proteins: Fcs and Sm‐fretmentioning

confidence: 99%

Single-molecule Spectroscopy: Exploring Heterogeneity in Chemical and Biological Systems

Ghosh

Bhattacharyya

2016

The Chemical Record

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Many chemical and biological systems are heterogeneous in the molecular length scale (∼ 1 nm). Heterogeneity in many chemical systems and organized assemblies may be monitored using single-molecule spectroscopy (SMS). In SMS, the size of the focal spot (i.e., the smallest region to be probed) is nearly half of the excitation wavelength (λ/2, i.e., 200-375 nm) for visible light (400-750 nm). We discuss how one can get spatial resolutions better than 200 nm using molecules as nanometric probes. We show that polymer hydrogels, lipid vesicles, room temperature ionic liquids (RTILs), and binary liquid mixtures exhibit such heterogeneity. Another important observation is solute-dependent friction in RTILs. In an RTIL, diffusion of an ionic solute is slower than that of a neutral solute.

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“…[3] Therefore, it is extremely important to understand the structural dynamics of cell-cell adhesion. Structural fluctuation is crucial in enzymes, [6][7][8][9][10][11][12] microtubule dynamics, [13] antigen-antibody complex, [14] and different organelles of al ive cell. [15][16][17] As an adhered system comprises of at least two different cell membranes, independent labeling of each cell membrane requires as uitable pair of dyes.…”

Section: Introductionmentioning

confidence: 99%

Probing Deviation of Adhered Membrane Dynamics between Reconstituted Liposome and Cellular System

Mondal

Chowdhury

Nandi

et al. 2019

Chemistry An Asian Journal

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The dynamics of cell-cell adhesion are complicated due to complexities in cellular interactions and intra-membrane interactions. In the present work, we have reconstituted al iposome-based model system to mimic the cell-cell adhesion process. Our model liposome system consists of one fluorescein-taggeda nd one TRITC (tetramethyl-rhodamine isothiocyanate)-tagged liposome, adhered through biotinneutravidin interaction. We monitored the adhesion process in liposomes using Fçrster Resonance EnergyT ransfer (FRET) between fluorescein( donor) and TRITC (acceptor). Occurrence of FRET is confirmed by the decrease in donor lifetime as well as distinct rise time of the acceptorf luorescence. Interestingly,t he acceptor'se mission exhibits fluctuations in the range of % 3 AE 1s.T his may be attributed to structural oscillationsa ssociated in two adhered liposomesa rising from the flexible nature of biotin-neutravidin interaction. We have compared the dynamics in ac ell-mimicking liposome system with that in an in vitro live cell system.I nt he adhered live cell system, we used CPM (7-diethylamino-3-(4maleimido-phenyl)-4-methylcoumarin, donor) andn ile red (acceptor), whicha re known to stain the membrane of CHO (Chinese Hamster Ovary) cells. The dynamicso ft he adhered membranes of two live CHO cells were observed through FRET between CPM and nile red. The acceptorf luorescence intensity exhibits an oscillation in the time-scaleo f% 1 AE 0.75 s, which is faster comparedt ot he reconstituted liposome system,i ndicating the contributions and involvement of multiple dynamic protein complexes around the cell membrane. This study offers simple reconstituted model systems to understand the complex membrane dynamics using aF RET-based physical chemistry approach.[a] Dr.

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Direct observation of the growth and shrinkage of microtubules by single molecule Förster resonance energy transfer

Cited by 12 publications

References 44 publications

Single-molecule fluorescence resonance energy transfer in molecular biology

Single-molecule fluorescence resonance energy transfer in molecular biology

Single-molecule Spectroscopy: Exploring Heterogeneity in Chemical and Biological Systems

Probing Deviation of Adhered Membrane Dynamics between Reconstituted Liposome and Cellular System

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