Cryo-electron microscopy as an investigative tool: the ribosome as an example.
Cryo-electron microscopy allows the visualization of macromolecules in their native state. Combined with techniques of three-dimensional reconstruction, cryo-EM images of single molecules can be used to study macromolecular interactions. The ribosome, a large RNA-protein complex with multiple binding interactions, is an excellent test case illustrating the power of these new techniques. Conformational changes during the binding of tRNA and protein factors to the ribosome can now be studied without the interference of crystal packing. Now that the first X-ray structures of ribosomal subunits have become available, conformational changes observed by cryo-EM in different functional states can be traced back to internal rearrangements of the underlying structural framework. Electron microscopy, X-ray crystallography, and modeling should be used together in the endeavor to understand the functioning of the translational machinery.
Frank
J
J
Howard Hughes Medical Institute, Health Research, Inc. at the Wadsworth Center, Empire State Plaza, Albany, New York 12201-0509, USA.
D000818:Animals; D020285:Cryoelectron Microscopy / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry / Q000648:ultrastructure; D046911:Macromolecular Substances; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D012333:RNA, Messenger / Q000737:chemistry / Q000648:ultrastructure; D012343:RNA, Transfer / Q000737:chemistry / Q000648:ultrastructure; D012269:Ribosomal Proteins / Q000737:chemistry / Q000648:ultrastructure; D012270:Ribosomes / Q000737:chemistry / Q000648:ultrastructure*
D046911:Macromolecular Substances; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D012343:RNA, Transfer