Electron crystallography of macromolecular periodic arrays on phospholipid monolayers.
Electron crystallography has the potential of yielding structural information equivalent to x-ray diffraction. The major difficulty has been preparing specimens with the required structural order and size for diffraction and imaging in the electron microscope. 2D crystallization on phospholipid monolayers is capable of fulfilling both of these requirements. Crystals can form as a result of specific interactions with a protein's ligand or an analog, suitably linked to a lipid tail; or on a surface of complementary head-group charge. With such choices, the availability of a suitable lipid is limited only by synthetic chemistry. Ultimately, it is the quality and regularity of the protein-protein interactions that determine the crystalline order, as it is with any protein crystal. In the case of streptavidin, the monolayer crystal diffracts beyond 2.5 A. A 3 A projection map reconstructed from electron diffraction amplitudes and phases from images shows density which can be interpreted as beta-sheets and clusters of side chains. It remains to be shown that the monolayer crystals are flat and diffract as well at high tilt angle as untilted. Technological issues such as charging must be resolved. With parallel advances in data collection and processing, electron crystallography of monolayer macromolecular crystals will eventually take its place beside x-ray crystallography and NMR as a routine and efficient structural technique.
Chiu
W
W
W.M. Keck Center for Computational Biology, Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, TX 77030, USA.
Avila-Sakar
A J
AJ
Schmid
M F
MF
D003461:Crystallography / Q000379:methods; D004798:Enzymes / Q000737:chemistry / Q000648:ultrastructure; D005615:Freezing; D008081:Liposomes; D008854:Microscopy, Electron / Q000379:methods; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000648:ultrastructure; D055672:Static Electricity
D004798:Enzymes; D008081:Liposomes; D011506:Proteins