Analysis of intracellular enzyme activity by surface enhanced Raman scattering

Ross Stevenson, Sarah McAughtrie, Laura Senior, Robert J Stokes, Helen McGachy, Laurence Tetley, Paola Nativo, James M Brewer, James Alexander, Karen Faulds, Duncan Graham

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Dysfunctional intracellular enzymatic activity is believed to be an underlying cause of a myriad of diseases. We present the first use of surface enhanced Raman scattering (SERS) as a detection technique capable of reporting intracellular activity of a specific enzyme. Careful choice of reagents allowed the preparation of high resolution cellular activity maps highlighting the specific conversion of the commonly used ELISA reagent 5-bromo-4-chloro-3-indolyl β-d-galactopyranoside (X-Gal), by wild type β-galactosidase enzymes. Further, through co-addition of X-Gal substrate and inhibitors we were able to demonstrate that intracellular substrate conversion occurred predominantly through an enzymatically specific pathway. The data presented therefore supports the application of SERS probes as sensitive, specific sensors of biochemical activity and demonstrates the use of SERS probes for the first time as beacons capable of high resolution subcellular localisation of native enzymes.
LanguageEnglish
Pages6331-6336
Number of pages6
JournalAnalyst
Volume138
Issue number21
Early online date29 Aug 2013
DOIs
Publication statusPublished - 7 Nov 2013

Fingerprint

Raman Spectrum Analysis
Enzyme activity
enzyme activity
Raman scattering
Enzymes
scattering
enzyme
probe
substrate
Substrates
beta-Galactosidase
Galactose
inhibitor
Enzyme-Linked Immunosorbent Assay
sensor
Sensors
analysis
5-bromo-4-chloro-3-indolyl beta-galactoside

Keywords

  • enzymatic activity
  • surface enhanced Raman scattering
  • SERS

Cite this

Stevenson, R., McAughtrie, S., Senior, L., Stokes, R. J., McGachy, H., Tetley, L., ... Graham, D. (2013). Analysis of intracellular enzyme activity by surface enhanced Raman scattering. Analyst, 138(21), 6331-6336. https://doi.org/10.1039/c3an00729d
Stevenson, Ross ; McAughtrie, Sarah ; Senior, Laura ; Stokes, Robert J ; McGachy, Helen ; Tetley, Laurence ; Nativo, Paola ; Brewer, James M ; Alexander, James ; Faulds, Karen ; Graham, Duncan. / Analysis of intracellular enzyme activity by surface enhanced Raman scattering. In: Analyst. 2013 ; Vol. 138, No. 21. pp. 6331-6336.
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Stevenson, R, McAughtrie, S, Senior, L, Stokes, RJ, McGachy, H, Tetley, L, Nativo, P, Brewer, JM, Alexander, J, Faulds, K & Graham, D 2013, 'Analysis of intracellular enzyme activity by surface enhanced Raman scattering' Analyst, vol. 138, no. 21, pp. 6331-6336. https://doi.org/10.1039/c3an00729d

Analysis of intracellular enzyme activity by surface enhanced Raman scattering. / Stevenson, Ross; McAughtrie, Sarah; Senior, Laura; Stokes, Robert J; McGachy, Helen; Tetley, Laurence; Nativo, Paola; Brewer, James M; Alexander, James; Faulds, Karen; Graham, Duncan.

In: Analyst, Vol. 138, No. 21, 07.11.2013, p. 6331-6336.

Research output: Contribution to journalArticle

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Stevenson R, McAughtrie S, Senior L, Stokes RJ, McGachy H, Tetley L et al. Analysis of intracellular enzyme activity by surface enhanced Raman scattering. Analyst. 2013 Nov 7;138(21):6331-6336. https://doi.org/10.1039/c3an00729d