Biopolym. Cell. 1987; 3(3):163-166.
Short Communications
Isolation of individual proteins from 50S subparticles of Thermus thermophilus ribosomes. crystallization of TL7 protein
1Sedelnikova S. E.
  1. Institute of Protein Research, Academy of Sciences of the USSR
    Pushchino, Moscow Region, USSR

Abstract

A method of selective extraction is suggested for eight proteins of 50S subparticles of ribosomes from extremal-thermophilic bacterium Thermus thermophilus at 60 °C in the presence of 0.5 M and 1 M NH4C1 and 50 % ethanol. As a result of chromatographic separation of extracted proteins on CM-sepharose CL five individual proteins (the purity is not less than 95 %) have been obtained. One of them, TL7 (probably, an analog of L6 protein from E. coli ribosomes), has been crystallized by the method of «hanging» drop with ammonium sulfate as a precipitant.

References

[1] Appelt K, Dijk J, Reinhardt R, Sanhuesa S, White SW, Wilson KS, Yonath A. The crystallization of ribosomal proteins from the 50 S subunit of the Escherichia coli and Bacillus stearothermophilus ribosome. J Biol Chem. 1981;256(22):11787-90.
[2] Appelt K, Dijk J, White SW, Wilson KS. Proteins of the Bacillus stearothermophilus ribosome. Crystallization of protein L6. FEBS Lett. 1983;160(1-2):75-7.
[3] Liljas A, Newcomer ME. Purification and crystallization of protein complex from Bacillus stearothermophilus ribosomes. J Mol Biol. 1981;153(2):393-8.
[4] Yonath A, Saper MA, Makowski I, Müssig J, Piefke J, Bartunik HD, Bartels KS, Wittmann HG. Characterization of single crystals of the large ribosomal particles from Bacillus stearothermophilus. J Mol Biol. 1986;187(4):633-6.
[5] Morikawa K, Kawakami M, Takemura S. Crystallization and preliminary X-ray diffraction study of 5 S rRNA from Thermus thermophilus HB8. FEBS Lett. 1982;145(2):194-6.
[6] Reshetnikova LS, Garber MB. Crystallization of elongation factor G from an extreme thermophile Thermus thermophilus HB8. FEBS Lett. 1983; 154(2):149-150.
[7] Karpova EA, Serdiuk IN, TarkhovskiÄ­ IuS, Orlova EV, Boroviagin VL. Crystallization of ribosomes from Thermus thermophilus. Dokl Akad Nauk SSSR. 1986;289(5):1263-6.
[8] Gogiya ZV, Yusupov MM, Spirina TN. Structure of Thermus thermophilus ribosomes. 1. Method of isolation and purification of ribosomes. Mol Biol (Mosk). 1986; 20(2):519-526.
[9] Hamel E, Koka M, Nakamoto T. Requirement of an Escherichia coli 50 S ribosomal protein component for effective interaction of the ribosome with T and G factors and with guanosine triphosphate. J Biol Chem. 1972;247(3):805-14.
[10] Kenny JW, Lambert JM, Traut RR. Cross-linking of ribosomes using 2-iminothiolane (methyl 4-mercaptobutyrimidate) and identification of cross-linked proteins by diagonal polyacrylamide/sodium dodecyl sulfate gel electrophoresis. Methods Enzymol. 1979;59:534-50.
[11] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[12] Tumanova LG, Gongadze GM, Gudkov AT. Isolation of all proteins from the 50S subparticles of ribosomes of Escherichia coli. Mol Biol (Mosk). 1982;16(4):807-13.