title: Electron microscopy of subnanometer surface features on metal-decorated protein crystals.
abstract: Crystals of heavy riboflavin synthase from Bacillus subtilis were freeze-etched and vacuum-coated at normal incidence with 0.1 to 0.4 nm of gold and silver, respectively. This decoration technique was applied to probe the protein surface for preferential nucleation sites. Image processing of the electron micrographs revealed two particular decoration sites for silver and a different one for gold. According to X-ray crystallography, the riboflavin synthase molecules are spherical and smooth except for a surface corrugation of less than 1 nm, which can not be depicted by heavy-metal shadowing. Thus the decoration sites represent sites of specific physical-chemical interactions between the condensing metal and the protein. The decoration pattern correctly reflects the icosahedral symmetry of the almost spherical protein molecules. Owing to the molecule's symmetry, the position of these topochemical sites with respect to the symmetry axes can be localized within 5A. The packing of the molecules in the crystal can be directly observed on shadowed replicas. Only decoration, however, makes it possible to observe the exact orientation of the molecules within the crystal planes and to derive the true lattice constant along the 6-fold screw axis. This proves decoration to be a technique suitable for studying crystal packing and the molecular symmetry of protein complexes at high resolution. The technique can be applied to crystals that are not large enough or insufficiently ordered for X-ray crystallography.
authors.affiliation: Institut für Technische Chemie, Technische Universität München, Garching, German Federal Republic.
authors.forename: L
authors.initials: L
authors.lastname: Bachmann
authors.forename: S
authors.initials: S
authors.lastname: Weinkauf
authors.forename: W
authors.initials: W
authors.lastname: Baumeister
authors.forename: I
authors.initials: I
authors.lastname: Wildhaber
authors.forename: A
authors.initials: A
authors.lastname: Bacher
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology; D003461:Crystallography; D005615:Freezing; D006046:Gold; D008854:Microscopy, Electron; D008961:Models, Structural; D012258:Riboflavin Synthase; D012834:Silver; D014166:Transferases
chemical_list: D012834:Silver; D006046:Gold; D014166:Transferases; D012258:Riboflavin Synthase