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

Record Type:	Language materials, manuscript : Monograph/item
Title/Author:	Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation./
Author:	Xu, Fang.
Description:	1 online resource (122 pages)
Notes:	Source: Dissertation Abstracts International, Volume: 79-04(E), Section: B.
Contained By:	Dissertation Abstracts International79-04B(E).
Subject:	Physical chemistry. -
Online resource:	click for full text (PQDT)
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. - 1 online resource (122 pages)

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

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

Includes bibliographical references

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.

Electronic reproduction.
Ann Arbor, Mich. :
ProQuest,
2018

Mode of access: World Wide Web

ISBN: 9780355539929Subjects--Topical Terms:

1148725
Physical chemistry.
Index Terms--Genre/Form:

554714
Electronic books.

Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation.
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245 1 0 $a Resonance-Energy-Transfer-Based Fluorescence Imaging and Free Energy Perturbation Calculation.
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300 $a 1 online resource (122 pages)
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500 $a Source: Dissertation Abstracts International, Volume: 79-04(E), Section: B.
500 $a Adviser: Richard A. Friesner.
502 $a Thesis (Ph.D.)--Columbia University, 2018.
504 $a Includes bibliographical references
520 $a 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.
533 $a Electronic reproduction. $b Ann Arbor, Mich. : $c ProQuest, $d 2018
538 $a Mode of access: World Wide Web
650 4 $a Physical chemistry. $3 1148725
655 7 $a Electronic books. $2 local $3 554714
690 $a 0494
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710 2 $a Columbia University. $b Chemistry. $3 1179813
773 0 $t Dissertation Abstracts International $g 79-04B(E).
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10622007 $z click for full text (PQDT)