Applications of electron cryo-microscopy to cardiovascular research.
Electron cryo-microscopy (cryo-EM) techniques have wide applications for the study of biological structures. The focus of this chapter is the use of cryo-EM and associated methods for the analysis of the three-dimensional (3D) structure of proteins and multicomponent macromolecular assemblies. Data evolving from these methods pertaining to the quaternary organization of proteins and protein-protein interactions, bridges an important gap between linear genomic information and understanding physiological function. This chapter provides general methods for examining two-dimensional crystalline arrays of proteins as well as single, randomly orientated protein complexes. It is significant that single-particle analysis of electron microscopy images has provided the only 3D data to-date for the two principal components of muscle excitation contraction coupling in the heart, namely the L-type voltage-gated calcium channel and the ryanodine receptor. This chapter describes approaches for identifying the extracellular and intracellular domains of the 3D structure of the L-type voltage-gated calcium channel and also incorporates general details for labeling and visualizing His-tagged proteins.
Kitmitto
Ashraf
A
School of Medicine, University of Manchester, UK.
D020746:Calcium Channels, L-Type / Q000737:chemistry; D002318:Cardiovascular Diseases / Q000473:pathology; D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D013194:Staining and Labeling / Q000379:methods
D020746:Calcium Channels, L-Type; D011506:Proteins