Abstract
| Language | English |
|---|---|
| Pages | 6331-6336 |
| Number of pages | 6 |
| Journal | Analyst |
| Volume | 138 |
| Issue number | 21 |
| Early online date | 29 Aug 2013 |
| DOIs | |
| Publication status | Published - 7 Nov 2013 |
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Keywords
- enzymatic activity
- surface enhanced Raman scattering
- SERS
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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 journal › Article
TY - JOUR
T1 - Analysis of intracellular enzyme activity by surface enhanced Raman scattering
AU - Stevenson, Ross
AU - McAughtrie, Sarah
AU - Senior, Laura
AU - Stokes, Robert J
AU - McGachy, Helen
AU - Tetley, Laurence
AU - Nativo, Paola
AU - Brewer, James M
AU - Alexander, James
AU - Faulds, Karen
AU - Graham, Duncan
PY - 2013/11/7
Y1 - 2013/11/7
N2 - 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.
AB - 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.
KW - enzymatic activity
KW - surface enhanced Raman scattering
KW - SERS
UR - http://www.scopus.com/inward/record.url?scp=84885000015&partnerID=8YFLogxK
U2 - 10.1039/c3an00729d
DO - 10.1039/c3an00729d
M3 - Article
VL - 138
SP - 6331
EP - 6336
JO - Analyst
T2 - Analyst
JF - Analyst
SN - 0003-2654
IS - 21
ER -