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Resonance-Energy-Transfer-Based Fluo...

Xu, Fang.

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Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation./
Author:	Xu, Fang.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2018,
Description:	122 p.
Notes:	Source: Dissertation Abstracts International, Volume: 79-04(E), Section: B.
Contained By:	Dissertation Abstracts International79-04B(E).
Subject:	Physical chemistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10622007
ISBN:	9780355539929

Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation.
Xu, Fang.

Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation. - Ann Arbor : ProQuest Dissertations & Theses, 2018 - 122 p.

Source: Dissertation Abstracts International, Volume: 79-04(E), Section: B.

Thesis (Ph.D.)--Columbia University, 2018.

This thesis focuses on an important aspect of protein functionality -- protein-protein interactions (PPI). Three physical chemistry techniques for or derived from protein-protein interaction investigation are discussed. First, in Chapter 2, we demonstrate a new fluorescent imaging technique that creates high-order nonlinear signals by harnessing the frustrated fluorescence resonance energy transfer (FRET) -- energy transfer between certain proteins close in proximity which is commonly used in PPI studies. In Chapter 3, we combine fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET), two most commonly used approaches to monitor protein-protein interactions in vivo, to create a novel hybrid strategy, bioluminescence assisted switching and fluorescence imaging (BASFI), which integrates the advantages of FRET and BRET. We demonstrate BASFI with Dronpa-RLuc8 fusion constructs and drug-inducible intermolecular FKBP-FRB protein-protein interactions in live cells with high sensitivity, resolution, and specificity. Finally, in Chapter 4, we propose a systematic free energy perturbation (FEP) protocol to computationally calculate the binding affinities between proteins. We demonstrate our protocol with the gp120 envelope glycoprotein of HIV-1 and three broadly neutralizing antibodies (bNAbs) of the VRC01 class and analyze antibody residues' contributions to the binding which further provides insights for antibody design.

ISBN: 9780355539929Subjects--Topical Terms:

1981412
Physical chemistry.

Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation.
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