Connect

facebook-icon linkedin-icon twitter-logo YouTube-icon make-a-gift-button-large


 

Full Stream Name: Supramolecular Sensors

Research Educator: Diana Zamora-Olivares

Principal Investigator: Eric V. Anslyn

Credit Options: Spring & Fall

In the Supramolecular Sensors Stream, students take on the challenge of constructing differential array sensors that can be ultimately applied to many fields such as homeland security, food industry, environmental science, and clinical diagnostics. The current project of the whole stream is the construction of peptide-based sensors that can be used to differentiate two types of samples, wine varietals, and protein kinases. The wine sensors work via the indicator displacement assay, a technique pioneered by Dr. Eric Anslyn and his research group at the Department of Chemistry and Biochemistry. The kinase biosensors work via a fluorescence-enhanced mechanism that is displayed when the biosensor is recognized by the protein kinase. 

 

fig1

(Photo credit: http://www.theage.com.au/news/epicure/out-with-the-old/2005/08/01/112274...)

 

Why Analyze Wine?

Wine is a perfect analyte to test the capability of differential sensors constructed due to the wine's chemical diversity and complexity. Compounds present in wine are metabolites coming not only from the grape but also from the yeast that act upon the grape, the environmental factors to which the growing grapes were exposed to, and even the oak barrels where the wine was aged. Wine is a great test sample for the results of the stream to extend to applications in disease diagnostics via metabolomic studies. It also translates to how sophisticated the sensor array is, even if the sensors constructed were relatively simple. Furthermore, there will be a direct impact of this study on the wine industry.

fig2

 

 

How will sensing be done?

Individual sensors function via indicator displacement, causing a change in signal, such as absorbance that can be measured using a UV-vis spectrophotometer. Using an array of receptors that are cross-reactive but not specific to analytes tested, a unique pattern of signals can be obtained from a specific analyte or analyte mixtures. Fingerprinting is facilitated using multivariate data analysis or pattern recognition protocols.

To learn more about this, contact Diana or Eric (see contact info below).

Fig3

Indicator Displacement Assay

fig4

 

Array sensing

 

Why analyze protein kinases?

There are about 500 different kinases in the Human Kinome. Particularly, the family of mitogen-activated protein kinases (MAPKs) has been involved in the development of 30% of all known cancers. The development of novel detection methods can allow us to understand the activation/inhibition of specific kinase pathways. MAPKs are very challenging proteins to recognize due to their similar structure.  The use of differential sensing techniques can be suitable for the detection and classification of these proteins not only in healthy cells, but also in cancerous cells. This research has the potential to impact targeted kinase therapies in the diagnosis of early-onset cancer.

 fig5

Mitogen-Activated Protein Kinases

How will sensing be done?

Individual biosensors function , causing an increase in the fluorescence intensity, which can be measured using a well plate reader. Using an array of peptide-based biosensors that are cross-reactive but not specific to specific kinases tested, a unique pattern of signals can be obtained from a particular kinase or mixtures of kinases such as in cell lysates. Fingerprinting is facilitated using multivariate data analysis, and quantification of kinases is measured with multivariate regression analysis such as support vector machines.

fig6

The fluorescence intensity of the peptide-based biosensor increases upon phosphorylation by a MAPK.

fig7

Array of biosensors showing differentiation of MAPKs

Moving Forward……

In order to expand the peptide repertoire we use to construct our sensors, the stream will be modifying the structure of peptides. Structural modifications on the peptides-based sensors can enhance the binding affinities towards its corresponding target, and thereby improve the detection of tannins in wines, and kinases in cell extracts respectively. This technology has been successfully used to study protein-protein and protein-drug interactions.  

What do you need to have to join and enjoy being in the stream?

A very important characteristic to have in the stream is an open mind. It is assumed that students have minimum knowledge about techniques and theories needed to perform and understand research in the stream. A high degree of patience is also great to have.

Some of the major techniques students will learn from the stream are:

 

  • Indicator displacement assay
  • Synthesis of biosensors
  • UV-Vis spectroscopy
  • Solid phase peptide synthesis
  • High performance liquid chromatography
  • High throughput techniques
  • Kinases assays
  • Multivariate data analysis

 

Contact info

Research Educator: Dr. Diana Zamora-Olivares  (click to email) 

WEL 2.102 (lab) and WEL 2.114 (office)

Principal Investigator: Dr. Eric V. Anslyn (click to email)  (512)-471-0068

Chemistry, Biochemistry Department Website: http://www.cm.utexas.edu/ 

 

 

Yes Yes
Biochemistry, Chemistry