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A new trend to determine biochemical parameters by quantitative FRET assays.

Liao JY, Song Y, Liu Y - Acta Pharmacol. Sin. (2015)

Bottom Line: Historically, FRET assays have been used to quantitatively deduce molecular distances.In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m).We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of California at Riverside, 900 University Avenue, Riverside, CA 92521, USA.

ABSTRACT
Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1-10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m). We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.

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Related in: MedlinePlus

Michaelis-Menten graphical analysis of CyPet–(pre-SUMO1)–YPet's processing by SENP1c. Data were plotted and analyzed by GraphPad Prism 5 of nonlinear regression.
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fig6: Michaelis-Menten graphical analysis of CyPet–(pre-SUMO1)–YPet's processing by SENP1c. Data were plotted and analyzed by GraphPad Prism 5 of nonlinear regression.

Mentions: The value of kcat can be directly calculated by dividing the experimentally determined value of vmax by the enzyme concentration, [E]. The Michaelis–Menten graph for the data in Table 3 is plotted in Figure 6.


A new trend to determine biochemical parameters by quantitative FRET assays.

Liao JY, Song Y, Liu Y - Acta Pharmacol. Sin. (2015)

Michaelis-Menten graphical analysis of CyPet–(pre-SUMO1)–YPet's processing by SENP1c. Data were plotted and analyzed by GraphPad Prism 5 of nonlinear regression.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4816234&req=5

fig6: Michaelis-Menten graphical analysis of CyPet–(pre-SUMO1)–YPet's processing by SENP1c. Data were plotted and analyzed by GraphPad Prism 5 of nonlinear regression.
Mentions: The value of kcat can be directly calculated by dividing the experimentally determined value of vmax by the enzyme concentration, [E]. The Michaelis–Menten graph for the data in Table 3 is plotted in Figure 6.

Bottom Line: Historically, FRET assays have been used to quantitatively deduce molecular distances.In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m).We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, University of California at Riverside, 900 University Avenue, Riverside, CA 92521, USA.

ABSTRACT
Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1-10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m). We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.

Show MeSH
Related in: MedlinePlus