Investigation of enzyme activity by SERRS using poly-functionalised benzotriazole derivatives as enzyme substrates

A.M. Ingram, Kirsten Stirling, K. Faulds, B.D. Moore, D. Graham

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

New methods of measuring biologically relevant concentrations of enzymes are necessary to allow greater understanding of biological systems. We have previously shown that aryl azo benzotriazolyl alkyl esters can act as enzyme substrates, with the progress of the reaction being monitored using SERRS (see Nat. Biotechnol., 2004, 22, 1133, ref. ). This is a wholly novel analytical application of SERRS, and the low detection levels of the technique allow for an ultra-sensitive enzyme assay. Masked enzyme substrates are used that are invisible to SERRS until enzymatic hydrolysis. Turnover of the substrate by the enzyme leads to the release of the surface-seeking dye necessary for SERRS, and intense signals are produced. Here we report an improved synthesis of 2H-benzotriazolyl alkyl esters via nucleophilic substitution of a chloromethyl ester by benzotriazolyl azo dyes, giving up to a ten-fold increase on previously reported yields. Introduction of electron-withdrawing groups to the benzotriazole ring allows control over the SERRS properties of the compounds. This is of great significance in expanding the synthetic flexibility and subsequently the fundamental use of these compounds as ultra-sensitive and selective reporters of enzyme activity.
LanguageEnglish
Pages2869-2873
Number of pages4
JournalOrganic and Biomolecular Chemistry
Volume4
Issue number15
DOIs
Publication statusPublished - 7 Aug 2006

Fingerprint

enzyme activity
Enzyme activity
enzymes
Derivatives
Substrates
Enzymes
esters
Esters
Azo Compounds
dyes
Enzymatic hydrolysis
Enzyme Assays
Biological systems
hydrolysis
Assays
flexibility
Hydrolysis
Substitution reactions
Coloring Agents
benzotriazole

Keywords

  • resonance raman scattering
  • SERRS
  • chemistry
  • alkylation
  • reagents
  • dyes

Cite this

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title = "Investigation of enzyme activity by SERRS using poly-functionalised benzotriazole derivatives as enzyme substrates",
abstract = "New methods of measuring biologically relevant concentrations of enzymes are necessary to allow greater understanding of biological systems. We have previously shown that aryl azo benzotriazolyl alkyl esters can act as enzyme substrates, with the progress of the reaction being monitored using SERRS (see Nat. Biotechnol., 2004, 22, 1133, ref. ). This is a wholly novel analytical application of SERRS, and the low detection levels of the technique allow for an ultra-sensitive enzyme assay. Masked enzyme substrates are used that are invisible to SERRS until enzymatic hydrolysis. Turnover of the substrate by the enzyme leads to the release of the surface-seeking dye necessary for SERRS, and intense signals are produced. Here we report an improved synthesis of 2H-benzotriazolyl alkyl esters via nucleophilic substitution of a chloromethyl ester by benzotriazolyl azo dyes, giving up to a ten-fold increase on previously reported yields. Introduction of electron-withdrawing groups to the benzotriazole ring allows control over the SERRS properties of the compounds. This is of great significance in expanding the synthetic flexibility and subsequently the fundamental use of these compounds as ultra-sensitive and selective reporters of enzyme activity.",
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Investigation of enzyme activity by SERRS using poly-functionalised benzotriazole derivatives as enzyme substrates. / Ingram, A.M.; Stirling, Kirsten; Faulds, K.; Moore, B.D.; Graham, D.

In: Organic and Biomolecular Chemistry, Vol. 4, No. 15, 07.08.2006, p. 2869-2873.

Research output: Contribution to journalArticle

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AU - Stirling, Kirsten

AU - Faulds, K.

AU - Moore, B.D.

AU - Graham, D.

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