Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

Fluorescent proteins of different colors are useful probes to study protein structure and function, and to investigate cellular events and conditions. Large efforts have focused on engineering new properties into fluorescent proteins via rational design and directed evolution. In addition to applications in imaging of protein expression level and subcellular localization, fluorescent proteins have been increasingly engineered to act as biosensors to track concentrations of small-molecule metabolites, enzyme activities, and protein conformational changes in living cells. Unlike small-molecule fluorescence biosensors, fluorescent proteins are genetically encodable, and thus can be expressed inside living cells. Attachment of organelle-specific signals to the proteins allows their localization to be specified. Recently, a new class of fluorescent protein biosensors has been developed to include unnatural amino acids as the sensing element. The unique chemical and physical properties of the unnatural amino acids enable sensor designs that cannot be realized by using the standard genetic code with the 20 canonical amino acids. In this chapter, we detail the general procedure for the genetic incorporation of unnatural amino acids. We further present two protocols for the in vitro and in vivo detection of hydrogen peroxide (H2O2) using a fluorescent protein biosensor that contains an unnatural amino acid, p-boronophenylalanine.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages191-219
Number of pages29
DOIs
StatePublished - Jan 1 2017

Publication series

NameMethods in Enzymology
Volume589
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Biosensing Techniques
Biosensors
Fluorescence
Amino Acids
Proteins
Cells
Genetic Code
Molecules
Enzyme activity
Metabolites
Organelles
Chemical properties
Hydrogen Peroxide
Color
Physical properties
Imaging techniques
Sensors

Keywords

  • Biosensors
  • Fluorescence
  • Fluorescent proteins
  • Genetic code expansion
  • Protein engineering
  • Unnatural amino acids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Niu, W., & Guo, J. (2017). Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids. In Methods in Enzymology (pp. 191-219). (Methods in Enzymology; Vol. 589). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2017.01.012

Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids. / Niu, Wei; Guo, Jiantao.

Methods in Enzymology. Academic Press Inc., 2017. p. 191-219 (Methods in Enzymology; Vol. 589).

Research output: Chapter in Book/Report/Conference proceedingChapter

Niu, W & Guo, J 2017, Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids. in Methods in Enzymology. Methods in Enzymology, vol. 589, Academic Press Inc., pp. 191-219. https://doi.org/10.1016/bs.mie.2017.01.012
Niu W, Guo J. Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids. In Methods in Enzymology. Academic Press Inc. 2017. p. 191-219. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2017.01.012
Niu, Wei ; Guo, Jiantao. / Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids. Methods in Enzymology. Academic Press Inc., 2017. pp. 191-219 (Methods in Enzymology).
@inbook{e1079204df2f4c41bf70a30592dfbeef,
title = "Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids",
abstract = "Fluorescent proteins of different colors are useful probes to study protein structure and function, and to investigate cellular events and conditions. Large efforts have focused on engineering new properties into fluorescent proteins via rational design and directed evolution. In addition to applications in imaging of protein expression level and subcellular localization, fluorescent proteins have been increasingly engineered to act as biosensors to track concentrations of small-molecule metabolites, enzyme activities, and protein conformational changes in living cells. Unlike small-molecule fluorescence biosensors, fluorescent proteins are genetically encodable, and thus can be expressed inside living cells. Attachment of organelle-specific signals to the proteins allows their localization to be specified. Recently, a new class of fluorescent protein biosensors has been developed to include unnatural amino acids as the sensing element. The unique chemical and physical properties of the unnatural amino acids enable sensor designs that cannot be realized by using the standard genetic code with the 20 canonical amino acids. In this chapter, we detail the general procedure for the genetic incorporation of unnatural amino acids. We further present two protocols for the in vitro and in vivo detection of hydrogen peroxide (H2O2) using a fluorescent protein biosensor that contains an unnatural amino acid, p-boronophenylalanine.",
keywords = "Biosensors, Fluorescence, Fluorescent proteins, Genetic code expansion, Protein engineering, Unnatural amino acids",
author = "Wei Niu and Jiantao Guo",
year = "2017",
month = "1",
day = "1",
doi = "10.1016/bs.mie.2017.01.012",
language = "English (US)",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "191--219",
booktitle = "Methods in Enzymology",

}

TY - CHAP

T1 - Novel Fluorescence-Based Biosensors Incorporating Unnatural Amino Acids

AU - Niu, Wei

AU - Guo, Jiantao

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Fluorescent proteins of different colors are useful probes to study protein structure and function, and to investigate cellular events and conditions. Large efforts have focused on engineering new properties into fluorescent proteins via rational design and directed evolution. In addition to applications in imaging of protein expression level and subcellular localization, fluorescent proteins have been increasingly engineered to act as biosensors to track concentrations of small-molecule metabolites, enzyme activities, and protein conformational changes in living cells. Unlike small-molecule fluorescence biosensors, fluorescent proteins are genetically encodable, and thus can be expressed inside living cells. Attachment of organelle-specific signals to the proteins allows their localization to be specified. Recently, a new class of fluorescent protein biosensors has been developed to include unnatural amino acids as the sensing element. The unique chemical and physical properties of the unnatural amino acids enable sensor designs that cannot be realized by using the standard genetic code with the 20 canonical amino acids. In this chapter, we detail the general procedure for the genetic incorporation of unnatural amino acids. We further present two protocols for the in vitro and in vivo detection of hydrogen peroxide (H2O2) using a fluorescent protein biosensor that contains an unnatural amino acid, p-boronophenylalanine.

AB - Fluorescent proteins of different colors are useful probes to study protein structure and function, and to investigate cellular events and conditions. Large efforts have focused on engineering new properties into fluorescent proteins via rational design and directed evolution. In addition to applications in imaging of protein expression level and subcellular localization, fluorescent proteins have been increasingly engineered to act as biosensors to track concentrations of small-molecule metabolites, enzyme activities, and protein conformational changes in living cells. Unlike small-molecule fluorescence biosensors, fluorescent proteins are genetically encodable, and thus can be expressed inside living cells. Attachment of organelle-specific signals to the proteins allows their localization to be specified. Recently, a new class of fluorescent protein biosensors has been developed to include unnatural amino acids as the sensing element. The unique chemical and physical properties of the unnatural amino acids enable sensor designs that cannot be realized by using the standard genetic code with the 20 canonical amino acids. In this chapter, we detail the general procedure for the genetic incorporation of unnatural amino acids. We further present two protocols for the in vitro and in vivo detection of hydrogen peroxide (H2O2) using a fluorescent protein biosensor that contains an unnatural amino acid, p-boronophenylalanine.

KW - Biosensors

KW - Fluorescence

KW - Fluorescent proteins

KW - Genetic code expansion

KW - Protein engineering

KW - Unnatural amino acids

UR - http://www.scopus.com/inward/record.url?scp=85013356507&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85013356507&partnerID=8YFLogxK

U2 - 10.1016/bs.mie.2017.01.012

DO - 10.1016/bs.mie.2017.01.012

M3 - Chapter

C2 - 28336064

AN - SCOPUS:85013356507

T3 - Methods in Enzymology

SP - 191

EP - 219

BT - Methods in Enzymology

PB - Academic Press Inc.

ER -