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Structural analysis of protein-RNA complexes by NMR spectroscopy (CAT#: STEM-MB-0685-WXH)

Introduction

The role of protein-RNA recognition is fundamental to many biological processes, protein-RNA interaction being at the heart of every molecular mechanisms controlling post-transcriptional gene expression. Deciphering the 3D structures of protein-RNA complexes is therefore of high significance. RNA binding proteins are very abundant in all kingdoms of life and often embed one to several small RNA binding domains. Since these domains often act as independent units, NMR spectroscopy is ideally suited to study the structure and dynamics of such domains in complex with their RNA targets.




Principle

Nuclear magnetic resonance spectroscopy, most commonly known as NMR spectroscopy or magnetic resonance spectroscopy (MRS), is a spectroscopic technique to observe local magnetic fields around atomic nuclei. The sample is placed in a magnetic field and the NMR signal is produced by excitation of the nuclei sample with radio waves into nuclear magnetic resonance, which is detected with sensitive radio receivers. The intramolecular magnetic field around an atom in a molecule changes the resonance frequency, thus giving access to details of the electronic structure of a molecule and its individual functional groups. As the fields are unique or highly characteristic to individual compounds, in modern organic chemistry practice, NMR spectroscopy is the definitive method to identify monomolecular organic compounds. Biochemists use NMR to identify proteins and other complex molecules. Besides identification, NMR spectroscopy provides detailed information about the structure, dynamics, reaction state, and chemical environment of molecules.

Applications

Nuclear Magnetic Resonance (NMR) spectroscopy is an analytical chemistry technique used in quality control and research for determining the content and purity of a sample as well as its molecular structure.

Procedure

1. Place the sample in a static magnetic field.
2. Excite nuclei in the sample with a radio frequency pulse.
41. Measure the frequency of the signals emitted by the sample.

Materials

NMR spectrometer