Analysis Biomolecular Interactions of the Holoenzyme with Individual Compounds by BLI (CAT#: STEM-MB-0137-CJ)

Name: Analysis Biomolecular Interactions of the Holoenzyme with Individual Compounds by BLI
SKU: STEM-MB-0137-CJ
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Introduction

Holoenzyme is a complete, functional enzyme, which is catalytically active. Holoenzyme consists of an apoenzyme together with its cofactors. Holoenzyme contains all the subunits required for the functioning of an enzyme, e.g. DNA polymerase III, RNA polymerase. And Holoenzyme = Apoenzyme + Cofactor
Holoenzyme is also known as a conjugate enzyme. The apoenzyme is the protein part of the enzyme, which is enzymatically inactive without cofactors. A cofactor is the non-protein part of the holoenzyme, which is essential for its activity. Cofactors can be metal ions (Mg2+, Fe3+, Zn2+) or organic molecules called coenzymes.

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Principle Applications Procedure Materials

Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Pharmacology

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.