Novel procedures for derivatization of ribosomes for crystallographic studies.
Undecagold and tetrairidium clusters have been used for the preparation of heavy-metal derivatives of ribosomal particles, necessary for the evaluation of phases in the x-ray structure determination of these large particles. To obtain specific binding, monofunctional reagents of the clusters were prepared and were covalently bound to free sulfhydryl groups on the surface of the ribosome. In addition, a mutant of Bacillus stearothermophilus which lacks one ribosomal protein (BL11) was grown. The heavy-atom clusters were covalently bound to isolated protein BL11, and the modified protein was consequently reconstituted into the mutated ribosomal subunits. Crystallographic data have been collected from crystals of native particles, from the mutated ones, and from the iridium- and gold-derivatized subunits. All these crystal forms are isomorphous within the experimental error.
Weinstein
S
S
Department of Structural Chemistry, Weizmann Institute, Rehovot, Israel.
Jahn
W
W
Hansen
H
H
Wittmann
H G
HG
Yonath
A
A
D003460:Crystallization; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D001411:Geobacillus stearothermophilus / Q000235:genetics / Q000648:ultrastructure; D009154:Mutation; D012269:Ribosomal Proteins / Q000302:isolation & purification; D012270:Ribosomes / Q000648:ultrastructure*; D014961:X-Ray Diffraction / Q000379:methods
D012269:Ribosomal Proteins