Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling

Eszter Kozma, Ivana Nikić, Balázs R. Varga, Iker Valle Aramburu, Jun Hee Kang, Oliver T. Fackler, Edward A. Lemke, P. Kele

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Introduction of bioorthogonal functionalities (e.g., trans-cyclooctene-TCO) into a protein of interest by site-specific genetic encoding of non-canonical amino acids (ncAAs) creates uniquely targetable platforms for fluorescent labeling schemes in combination with tetrazine-functionalized dyes. However, fluorescent labeling of an intracellular protein is usually compromised by high background, arising from the hydrophobicity of ncAAs; this is typically compensated for by hours-long washout to remove excess ncAAs from the cellular interior. To overcome these problems, we designed, synthesized, and tested new, hydrophilic TCO-ncAAs. One derivative, DOTCO-lysine was genetically incorporated into proteins with good yield. The increased hydrophilicity shortened the excess ncAA washout time from hours to minutes, thus permitting rapid labeling and subsequent fluorescence microscopy.

Original languageEnglish
Pages (from-to)1518-1524
Number of pages7
JournalChemBioChem
DOIs
Publication statusPublished - Aug 17 2016

Fingerprint

Labeling
Amino Acids
Hydrophobic and Hydrophilic Interactions
Proteins
Fluorescence microscopy
Hydrophilicity
Hydrophobicity
Fluorescence Microscopy
Lysine
Coloring Agents
Derivatives

Keywords

  • amino acids
  • click chemistry
  • fluorescence
  • hydrophilicity
  • protein engineering

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

Cite this

Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling. / Kozma, Eszter; Nikić, Ivana; Varga, Balázs R.; Aramburu, Iker Valle; Kang, Jun Hee; Fackler, Oliver T.; Lemke, Edward A.; Kele, P.

In: ChemBioChem, 17.08.2016, p. 1518-1524.

Research output: Contribution to journalArticle

Kozma, E, Nikić, I, Varga, BR, Aramburu, IV, Kang, JH, Fackler, OT, Lemke, EA & Kele, P 2016, 'Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling', ChemBioChem, pp. 1518-1524. https://doi.org/10.1002/cbic.201600284
Kozma, Eszter ; Nikić, Ivana ; Varga, Balázs R. ; Aramburu, Iker Valle ; Kang, Jun Hee ; Fackler, Oliver T. ; Lemke, Edward A. ; Kele, P. / Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling. In: ChemBioChem. 2016 ; pp. 1518-1524.
@article{06d6c2936e394e289c843fa2a2ad9541,
title = "Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling",
abstract = "Introduction of bioorthogonal functionalities (e.g., trans-cyclooctene-TCO) into a protein of interest by site-specific genetic encoding of non-canonical amino acids (ncAAs) creates uniquely targetable platforms for fluorescent labeling schemes in combination with tetrazine-functionalized dyes. However, fluorescent labeling of an intracellular protein is usually compromised by high background, arising from the hydrophobicity of ncAAs; this is typically compensated for by hours-long washout to remove excess ncAAs from the cellular interior. To overcome these problems, we designed, synthesized, and tested new, hydrophilic TCO-ncAAs. One derivative, DOTCO-lysine was genetically incorporated into proteins with good yield. The increased hydrophilicity shortened the excess ncAA washout time from hours to minutes, thus permitting rapid labeling and subsequent fluorescence microscopy.",
keywords = "amino acids, click chemistry, fluorescence, hydrophilicity, protein engineering",
author = "Eszter Kozma and Ivana Nikić and Varga, {Bal{\'a}zs R.} and Aramburu, {Iker Valle} and Kang, {Jun Hee} and Fackler, {Oliver T.} and Lemke, {Edward A.} and P. Kele",
year = "2016",
month = "8",
day = "17",
doi = "10.1002/cbic.201600284",
language = "English",
pages = "1518--1524",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley-VCH Verlag",

}

TY - JOUR

T1 - Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling

AU - Kozma, Eszter

AU - Nikić, Ivana

AU - Varga, Balázs R.

AU - Aramburu, Iker Valle

AU - Kang, Jun Hee

AU - Fackler, Oliver T.

AU - Lemke, Edward A.

AU - Kele, P.

PY - 2016/8/17

Y1 - 2016/8/17

N2 - Introduction of bioorthogonal functionalities (e.g., trans-cyclooctene-TCO) into a protein of interest by site-specific genetic encoding of non-canonical amino acids (ncAAs) creates uniquely targetable platforms for fluorescent labeling schemes in combination with tetrazine-functionalized dyes. However, fluorescent labeling of an intracellular protein is usually compromised by high background, arising from the hydrophobicity of ncAAs; this is typically compensated for by hours-long washout to remove excess ncAAs from the cellular interior. To overcome these problems, we designed, synthesized, and tested new, hydrophilic TCO-ncAAs. One derivative, DOTCO-lysine was genetically incorporated into proteins with good yield. The increased hydrophilicity shortened the excess ncAA washout time from hours to minutes, thus permitting rapid labeling and subsequent fluorescence microscopy.

AB - Introduction of bioorthogonal functionalities (e.g., trans-cyclooctene-TCO) into a protein of interest by site-specific genetic encoding of non-canonical amino acids (ncAAs) creates uniquely targetable platforms for fluorescent labeling schemes in combination with tetrazine-functionalized dyes. However, fluorescent labeling of an intracellular protein is usually compromised by high background, arising from the hydrophobicity of ncAAs; this is typically compensated for by hours-long washout to remove excess ncAAs from the cellular interior. To overcome these problems, we designed, synthesized, and tested new, hydrophilic TCO-ncAAs. One derivative, DOTCO-lysine was genetically incorporated into proteins with good yield. The increased hydrophilicity shortened the excess ncAA washout time from hours to minutes, thus permitting rapid labeling and subsequent fluorescence microscopy.

KW - amino acids

KW - click chemistry

KW - fluorescence

KW - hydrophilicity

KW - protein engineering

UR - http://www.scopus.com/inward/record.url?scp=84982218124&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84982218124&partnerID=8YFLogxK

U2 - 10.1002/cbic.201600284

DO - 10.1002/cbic.201600284

M3 - Article

C2 - 27223658

AN - SCOPUS:84982218124

SP - 1518

EP - 1524

JO - ChemBioChem

JF - ChemBioChem

SN - 1439-4227

ER -