Analysis of intracellular enzyme activity by surface enhanced Raman scattering

Stevenson, Ross and McAughtrie, Sarah and Senior, Laura and Stokes, Robert J and McGachy, Helen and Tetley, Laurence and Nativo, Paola and Brewer, James M and Alexander, James and Faulds, Karen and Graham, Duncan (2013) Analysis of intracellular enzyme activity by surface enhanced Raman scattering. Analyst, 138 (21). pp. 6331-6336.

[img]
Preview
PDF (Stevenson-etalAnalyst2013-intracellular-enzyme-activity)
Stevenson_etalAnalyst2013_intracellular_enzyme_activity.pdf
Final Published Version
License: Creative Commons Attribution 4.0 logo

Download (621kB)| Preview

    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.