scholarly journals Two-dimensional Fluorescence Lifetime Correlation Spectroscopy on the Conformational Dynamics of Proteins

2014 ◽  
Vol 54 (6) ◽  
pp. 321-322
Author(s):  
Takuhiro OTOSU
Keyword(s):  
Fluorescence Lifetime ◽  
Conformational Dynamics ◽  
Correlation Spectroscopy ◽  
Two Dimensional

scholarly journals 3P052 Two-dimensional fluorescence lifetime correlation spectroscopy on the conformational dynamics of the unfolded state of BdpA(01A. Protein: Structure,Poster)

2013 ◽  
Vol 53 (supplement1-2) ◽  
pp. S220
Author(s):  
Takuhiro Otosu ◽  
Kunihiko Ishii ◽  
Hiroyuki Oikawa ◽  
Munehito Arai ◽  
Satoshi Takahashi ◽  
...  
Keyword(s):  
Protein Structure ◽  
Fluorescence Lifetime ◽  
Conformational Dynamics ◽  
Correlation Spectroscopy ◽  
Two Dimensional ◽  
Unfolded State

scholarly journals Two-Dimensional Fluorescence Lifetime Correlation Spectroscopy: Concepts and Applications

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2972 ◽  
Author(s):  
Takuhiro Otosu ◽  
Shoichi Yamaguchi
Keyword(s):  
Single Molecule ◽  
Fluorescence Lifetime ◽  
Fluorescence Correlation Spectroscopy ◽  
Conformational Dynamics ◽  
Correlation Spectroscopy ◽  
Two Dimensional ◽  
Single Molecule Level ◽  
Fluorescence Correlation ◽  
Promising Tool ◽  
Molecular Ruler

We review the basic concepts and recent applications of two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS), which is the extension of fluorescence correlation spectroscopy (FCS) to analyze the correlation of fluorescence lifetime in addition to fluorescence intensity. Fluorescence lifetime is sensitive to the microenvironment and can be a “molecular ruler” when combined with FRET. Utilization of fluorescence lifetime in 2D FLCS thus enables us to quantify the inhomogeneity of the system and the interconversion dynamics among different species with a higher time resolution than other single-molecule techniques. Recent applications of 2D FLCS to various biological systems demonstrate that 2D FLCS is a unique and promising tool to quantitatively analyze the microsecond conformational dynamics of macromolecules at the single-molecule level.

scholarly journals Two-Dimensional Fluorescence Lifetime Correlation Spectroscopy on the Folding Mechanism of B Domain of Protein A

2015 ◽  
Vol 108 (2) ◽  
pp. 501a
Author(s):  
Takuhiro Otosu ◽  
Kunihiko Ishii ◽  
Hiroyuki Oikawa ◽  
Munehito Arai ◽  
Satoshi Takahashi ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Protein A ◽  
Correlation Spectroscopy ◽  
Two Dimensional ◽  
Folding Mechanism

scholarly journals Fast and Robust 2D Inverse Laplace Transformation of Single-Molecule Fluorescence Lifetime Data

2021 ◽  
Author(s):  
Saurabh Talele ◽  
John T. King
Keyword(s):  
Nmr Spectroscopy ◽  
Fluorescence Spectroscopy ◽  
Single Molecule ◽  
Fluorescence Lifetime ◽  
Conformational Dynamics ◽  
Correlation Spectroscopy ◽  
Great Promise ◽  
Biological Macromolecules ◽  
Single Molecule Fluorescence ◽  
Laplace Transformations

AbstractFluorescence spectroscopy at the single-molecule scale has been indispensable for studying conformational dynamics and rare states of biological macromolecules. Single-molecule 2D-fluorescence lifetime correlation spectroscopy (sm-2D-FLCS) is an emerging technique that holds great promise for the study of protein and nucleic acid dynamics as it 1) resolves conformational dynamics using a single chromophore, 2) measures forward and reverse transitions independently, and 3) has a dynamic window ranging from microseconds to seconds. However, the calculation of a 2D fluorescence relaxation spectrum requires an inverse Laplace transition (ILT), which is an ill-conditioned inversion that must be estimated numerically through a regularized minimization. The current methods for performing ILTs of fluorescence relaxation can be computationally inefficient, sensitive to noise corruption, and difficult to implement. Here, we adopt an approach developed for NMR spectroscopy (T1-T2 relaxometry) to perform 1D and 2D-ILTs on single-molecule fluorescence spectroscopy data using singular-valued decomposition and Tikhonov regularization. This approach provides fast, robust, and easy to implement Laplace inversions of single-molecule fluorescence data.Significance StatementInverse Laplace transformations are a powerful approach for analyzing relaxation data. The inversion computes a relaxation rate spectrum from experimentally measured temporal relaxation, circumventing the need to choose appropriate fitting functions. They are routinely performed in NMR spectroscopy and are becoming increasing used in single-molecule fluorescence experiments. However, as Laplace inversions are ill-conditioned transformations, they must be estimated from regularization algorithms that are often computationally costly and difficult to implement. In this work, we adopt an algorithm first developed for NMR relaxometry to provide fast, robust, and easy to implement 1D and 2D inverse Laplace transformations on single-molecule fluorescence data.

Sign in / Sign up

Export Citation Format

Share Document