Bacillus stearothermophilus, ribosomal subunits

Our first microcrystals were of Bacillus stearothermophilus SOS subunits and of 70S ribosomes from E. coli they were mainly grown from lower alcohols, toluene, or chloroform. Each ribosomal preparation required slightly different crystallization conditions, and often the preparation had almost been exhausted by the time conditions were optimised. We also found that crystals grew from active particles only. 

Appelt, K., Dijk, J., Reinhardt, R., Sanhuesa, S., White, S.W., Wilson, K.S., and Yonath, A., 1981, The crystallization of ribosomal proteins from the SOS subunit of the Escherichia coli and Bacillus stearothermophilus ribosome, J. Biol. Chem., 256 11787. 

The only other E. coli ribosomal protein whose crystallization has so far been reported is L29 (Appelt et al., 1981). On the other hand, attempts to crystallize ribosomal proteins from the thermophilic Bacillus stearothermophilus have been more successful. Protein BL17, which according to its amino acid sequence (Kimura et al., 1980) corresponds to protein L9 from the E. coli ribosome (Kimura et al., 1982), was the first intact ribosomal protein to give crystals useful for X-ray structural analysis (Appelt et al., 1979). Several other B. stearothermophUus ribosomal proteins, namely BL6 and BL30 (Appelt eteU., 1981,1983) from the large and BS5 (Appelt et al., 1983) from the small subunit have been crystallized, and the determination of their three-dimensional structure at a resolution of better than 3 A is now in progress. Furthermore, crystals of aB. stearothermophilus ribosomal protein complex, which corresponds to the complex (L7/L12)4 LIO from E. coli ribosome, have been obtained (Liljas and Newcomer, 1981). 

Fig. 7. (a) Crystals of E. colt 70 S ribosomes, (b) and (c) Electron micrographs of sections through three-dimensional crystals shown in (a) in two orthogonal directions (Wittmann et al., 1982). (d) and (e) Crystals and computed filtered image of a section through a crystal of Bacillus stearothermophilus 50 S ribosomal subunits (Yonath et al., 1982a,b Leonard et al., 1983). (d) and (e) are related to two different crystal forms. Reproduced with permission from Wittmann (1983). 

Yonath, A.E., Mussig, J., Tesche, B., Lorenz, S., Erdmann, V.A., and Wittmann, H.G., 1980, Crystallization of the large ribosomal subunits from Bacillus stearothermophilus, B iochem. Internat., 1 428. 

---