Atomic force microscopy in structural biology: from the subcellular to the submolecular.
Atomic force microscopy (AFM) is capable of generating images within ranges of resolution that are of particular interest in biology. Although atomic resolution may not be possible with biological samples, a great deal of information can still be obtained from images that provide structures at a slightly lower level of resolution. The submolecular resolution images of bacteriorhodopsin and the chaperonin GroES, which revealed, respectively, individual loops and beta-turns, confirmed and complemented other structural investigations, while the molecular-level features in images of membrane-bound VacA, a cytotoxin from Helicobacter pylori, immediately suggested the possibility, subsequently proven, of channel-forming ability. A series of images with macromolecular resolution directly provided details on the mechanisms by which RNA polymerase nonspecifically translocates along DNA, and images with subcellular resolving power of erythrocytic cellular membranes showed, with unambiguous clarity, linear arrays of molecular complexes. In this review, we will describe some of the most biologically relevant findings that have been obtained with AFM within ranges of resolution from the submolecular to the molecular, and from the macromolecular to the subcellular. Furthermore, we will describe some of the sample conditions and imaging environments that are likely important to achieve a particular level of resolution.
Czajkowsky
D M
DM
Department of Molecular Physiology, University of Virginia School of Medicine, Charlottesville 22908, USA. dmc2m@virginia.edu
Iwamoto
H
H
Shao
Z
Z
D001426:Bacterial Proteins / Q000648:ultrastructure; D001427:Bacterial Toxins; D001695:Biology / Q000379:methods; D002477:Cells / Q000648:ultrastructure; D018835:Chaperonin 10 / Q000648:ultrastructure; D006460:Hemolysin Proteins; D007075:Immunoglobulin M / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D009124:Muscle Proteins / Q000648:ultrastructure
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D018835:Chaperonin 10; D006460:Hemolysin Proteins; D007075:Immunoglobulin M; D009124:Muscle Proteins; C074689:VacA protein, Helicobacter pylori; C021091:staphylococcal alpha-toxin