Question

Please describe the advantages of using cryo-electron microscopy in structure determination and the technology advancement that makes cryo-EM an attractive approach to resolve protein structure at atomic resolution.

Answer 1

Ans:

Advantages of using cryo-electron microscopy in structure determination

-Advantage of cryo-electron microscopy is that no crystallization of molecule of interest is required. Since many proteins/macromolecules can not be crystallised and there are macromolecules whose structures change irreversibly by crystallisation process.

-Proteins/macromolecules can be observed in all possible conformations.

-Cryo-electron microscopy samples are not needed to be dehydrated or stained, thus their structure remains intact. Therefore false shapes generated as a result of dehydration and staining is avoided in Cryo-EM.

-Frozen samples are less likely to be damaged by electron microscope.

Technology advancement that makes cryo-EM an attractive approach to resolve protein structure at atomic resolution

Cryo-electron microscopy of non-crystalline single particles is used to determine the structure of biological macromolecules. Technological afvances e.g development of electron detectors and effective computational image analysis techniques are revolutionizing uses of cryo-EM. In addition instrumentation and software have increased the potential of cryo-EM and help in productiojn of structures at resolutions allowing atomic modeling into density map.