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Host–pathogen interactions by SDSL EPR (CAT#: STEM-MB-1029-WXH)

Introduction

The interaction between a pathogen and its host represents the first step in infectious diseases and detailed information about this interaction down to the molecular level can provide means for the development of drugs that can effectively prevent infection, rendering this field of great importance in biomedicine. In general, such kind of drugs can tackle two different points of action: (i) it can prevent the interaction by competitive binding to one of the interaction partners – for this approach detailed knowledge about the structural nature of the interaction is required, or (ii) it can prevent synthesis and/or assembly of the components that comprise the interaction site of the pathogen – in this case information about the mechanisms of synthesis/assembly of this components is necessary.




Principle

Electron Paramagnetic Resonance (EPR), also called Electron Spin Resonance (ESR), is a branch of magnetic resonance spectroscopy which utilizes microwave radiation to probe species with unpaired electrons, such as radicals, radical cations, and triplets in the presence of an externally applied static magnetic field.
EPR spectroscopy is particularly suitable for the investigation of (bio)chemical systems with strongly localized spin density and their interaction with the environment. For these systems EPR provides information on the structure and dynamics and is widely used in chemistry, physics and biology.
Site-directed spin labeling (SDSL) is a technique for investigating the structure and local dynamics of proteins using electron spin resonance. The theory of SDSL is based on the specific reaction of spin labels with amino acids. A spin label's built-in protein structure can be detected by EPR spectroscopy. SDSL is also a useful tool in examinations of the protein folding process.

Applications

• Study dynamic organisation of lipids in biological membranes, lipid-protein interactions and temperature of transition of gel to liquid crystalline phases.
• Determine oxygen levels in tissues and blood.
• Injection of spin-labeled molecules allows for electron resonance imaging of living organisms.
• EPR can be used to measure microviscosity and micropolarity within drug delivery systems as well as the characterization of colloidal drug carriers.
• The study of radiation-induced free radicals in biological substances (for cancer research).
• Investigation on the antioxidant properties of medicine

Procedure

1. Sample Preparation
2. Electron paramagnetic resonance (EPR) spectroscopy testing
3. Data analysis

Materials

• EPR Spectrometer
• Spectrophotometer