Non-natural amino acid fluorophores for one- and two-step fluorescence resonance energy transfer applications

Rogers, Julie M.; Lippert, Lisa G.; Gai, Feng

doi:10.1016/j.ab.2009.12.027

Cited by 35 publications

(46 citation statements)

References 55 publications

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“…However, due to low QYs of both Phe and Tyr, Trp is the only widely used natural fluorescent probe. To overcome this limitation nonnatural (also called unnatural or noncanonical) amino acids have been fabricated [207][208][209][210][211][212][213]. These nonnatural amino acids can provide larger QYs and new FRET pairs for protein structure function analysis.…”

Section: Nonnatural Amino Acidsmentioning

confidence: 99%

“…The Trp-PheCN FRET pair has been used to determine detailed protein folding and unfolding in two small proteins, the villin headpiece subdomain (HP35) and the lysin motif (LysM) domain [200]. Two other nonnatural amino acids, 7-azatryptophan (7AW) and 5-hydroxytryptophan (5HW) ( Figure 6.13), were recently found to be FRET acceptors to PheCN [210]. The 7AW-PheCN FRET pair had a greater separation of fluorescent spectrums than PheCN-Trp.…”

Section: Nonnatural Amino Acidsmentioning

confidence: 99%

“…The 7AW-PheCN FRET pair had a greater separation of fluorescent spectrums than PheCN-Trp. Moreover PheCN, Trp, and 7AW can be used in a multistep FRET system to investigate interactions of three points on a protein (see Section 6.6) [210]. Even newer nonnatural FRET pairs are continuously being developed.…”

Section: Nonnatural Amino Acidsmentioning

confidence: 99%

“…Such systems have potential use in solar cells, nanoscale photonic devices, and other optoelectronic applications [269,567,568]. More commonly, these multi-FRET configurations are developed as a means to extend the optical ruler and are used to elucidate biological configurations, study protein and DNA interactions, and for biosensing applications [135,210,365,366,412,413,569].…”

Section: Multi-fret Systemsmentioning

confidence: 99%

“…Copyright 2005, American Chemical Society.) urea-induced conformational changes [210]. The nonnatural amino acid fluorophores, PheCN and 7AW (Section 6.4.2), were incorporated by mutation of the Cand N-terminus.…”

Section: Multi-fret Systemsmentioning

confidence: 99%

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Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

Sapsford

Wildt

Mariani

et al. 2013

FRET – Förster Resonance Energy Transfer

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The intrinsic sensitivity (r 6 dependence) of F€ orster (or fluorescence) resonance energy transfer (FRET) to nanoscale changes in the donor/acceptor separation distance has made FRET an invaluable biophysical tool in a variety of applications, ranging from studying the structure and conformation of proteins and nucleic acids to examining biomolecular interactions, including its use in in vitro and in vivo bioassays [1][2][3][4][5][6][7]. While the myriad of FRET configurations and techniques currently in use are covered throughout this book, here we focus primarily on the materials utilized as donor or acceptor probes in FRET rather than the process itself [3,5,8,9]. Our 2006 review paper on this topic serves as the foundation for this updated chapter [3]. The materials were divided into three main categories: organic materials that include "traditional" dye fluorophores, dark quenchers, polymers, and carbon nanomaterials (NMs); inorganic materials such as metal chelates, metal, and semiconductor nanocrystals; and fluorophores of biological origin such as fluorescent proteins (FPs), amino acids, and fluorescence generated from enzymatic bioluminescence (BL) and chemiluminescence (CL). These materials may function as FRET donors and/or acceptors, depending upon experimental design. Many of the new materials developed and/or new donor-acceptor probe combinations used address some of the inherent complications of more traditional FRET materials, including photobleaching, spectral cross talk, and direct excitation of the acceptor species, and examples of these will be highlighted and discussed throughout the chapter. Since the vast majority of FRET applications are biological in nature, they routinely involve some type of biomolecule labeling strategy, which ultimately plays a significant and fundamental role in the success and interpretation of the resulting FRET. Therefore, we begin the chapter with a brief discussion of the bioconjugation techniques commonly utilized for FRET and points to consider, followed by sections highlighting the current and emerging materials that are used, or have the potential to be used, in FRET applications.

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Section: Nonnatural Amino Acidsmentioning

confidence: 99%

Section: Nonnatural Amino Acidsmentioning

confidence: 99%

Section: Nonnatural Amino Acidsmentioning

confidence: 99%

Section: Multi-fret Systemsmentioning

confidence: 99%

Section: Multi-fret Systemsmentioning

confidence: 99%

See 3 more Smart Citations

Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

Sapsford

Wildt

Mariani

et al. 2013

FRET – Förster Resonance Energy Transfer

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Kinetics of peptide folding in lipid membranes

2015

Self Cite

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Despite our extensive understanding of water-soluble protein folding kinetics, much less is known about the folding dynamics and mechanisms of membrane proteins. However, recent studies have shown that for relatively simple systems, such as peptides that form a transmembrane α-helix, helical dimer, or helix-turn-helix, it is possible to assess the kinetics of several important steps, including peptide binding to the membrane from aqueous solution, peptide folding on the membrane surface, helix insertion into the membrane, and helix-helix association inside the membrane. Herein, we provide a brief review of these studies and also suggest new initiation and probing methods that could lead to improved temporal and structural resolution in future experiments.

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Folding stability modulation of the villin headpiece helical subdomain by 4‐fluorophenylalanine and 4‐methylphenylalanine

2015

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HP36, the helical subdomain of villin headpiece, contains a hydrophobic core composed of three phenylalanine residues (Phe47, Phe51, and Phe58). Hydrophobic effects and electrostatic interactions were shown to be the critical factors in stabilizing this core and the global structure. To assess the interactions among Phe47, Phe51, and Phe58 residues and investigate how they affect the folding stability, we implanted 4-fluorophenylalanine (Z) and 4-methylphenylalanine (X) into the hydrophobic core of HP36. We chemically synthesized HP36 and its seven variants including four single mutants whose Phe51 or Phe58 was replaced with Z or X, and three double mutants whose Phe51 and Phe58 were both substituted. Circular dichroism and nuclear magnetic resonance measurements show that the variants exhibit a native HP36 like fold, of which F51Z and three double mutants are more stable than the wild type. Molecular modeling provided detailed interaction energy within the phenylalanine residues, revealing that electrostatic interactions dominate the stability modulation upon the introduction of 4-fluorophenylalanine and 4-methylphenylalanine. Our results show that these two non-natural amino acids can successfully tune the interactions in a relatively compact hydrophobic core and the folding stability without inducing dramatic steric effects. Such an approach may be applied to other folded motifs or proteins.

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Non-natural amino acid fluorophores for one- and two-step fluorescence resonance energy transfer applications

Cited by 35 publications

References 55 publications

Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

Materials for FRET Analysis: Beyond Traditional Dye–Dye Combinations

Kinetics of peptide folding in lipid membranes

Folding stability modulation of the villin headpiece helical subdomain by 4‐fluorophenylalanine and 4‐methylphenylalanine

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