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Related Concept Videos

Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
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Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
Enzyme Kinetics01:19

Enzyme Kinetics

Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...

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A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1
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Supramolecular tandem enzyme assays.

Roy N Dsouza1, Andreas Hennig, Werner M Nau

  • 1School of Engineering and Science, Jacobs University Bremen, Campus Ring 1, 28759 Bremen, Germany.

Chemistry (Weinheim an Der Bergstrasse, Germany)
|February 28, 2012
PubMed
Summary

A new supramolecular tandem assay uses fluorescent dye binding to macrocyclic hosts for enzyme activity measurement. This innovative method enables rapid screening for enzyme inhibitors, activators, and enantiomeric excess determination.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Supramolecular Chemistry

Background:

  • Enzyme assays are crucial for drug discovery and diagnostics.
  • Current methods can be complex and time-consuming.
  • Need for rapid, adaptable assay technologies.

Purpose of the Study:

  • To introduce a novel enzyme assay methodology.
  • To demonstrate its versatility in various biochemical applications.
  • To facilitate high-throughput screening (HTS).

Main Methods:

  • Utilizing reversible, competitive binding of a fluorescent dye and enzyme reaction components (substrate/product) to a macrocyclic host.
  • Developing the
  • supramolecular tandem assay
  • platform.
  • Implementing fluorescence-based read-out.

Main Results:

  • Successful application of the assay for inhibitor and activator screening.
  • Demonstrated utility in sensor array development.
  • Effective determination of enantiomeric excess for amino acids.

Conclusions:

  • The supramolecular tandem assay offers a simple, rapid, and versatile approach for enzyme activity analysis.
  • Its fluorescence-based detection is ideal for high-throughput screening.
  • This method has broad applicability in biochemical research and diagnostics.