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Conformational Analysis of the ABC Transporter MsbA by Electron paramagnetic resonance (EPR) spectroscopy (CAT#: STEM-MB-1069-WXH)

Introduction

ATP-binding cassette (ABC) transporters are an example of ATP-dependent pumps. ABC transporters are ubiquitous membrane-bound proteins, present in all prokaryotes, as well as plants, fungi, yeast and animals. These pumps can move substrates in (influx) or out (efflux) of cells. MsbA is an essential ABC (ATP-binding cassette) transporter involved in lipid A transport across the cytoplasmic membrane of Gram-negative bacteria.




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.

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
33. Data analysis

Materials

• EPR Spectrometer
• Spectrophotometer