Isolation of individual proteins from 50S subparticles of Thermus thermophilus ribosomes. crystallization of TL7 protein

Authors

  • S. E. Sedelnikova Institute of Protein Research, Academy of Sciences of the USSR Pushchino, Moscow Region, USSR Author

DOI:

https://doi.org/10.7124/bc.0001EB

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

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.

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.

Liljas A, Newcomer ME. Purification and crystallization of protein complex from Bacillus stearothermophilus ribosomes. J Mol Biol. 1981;153(2):393-8.

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.

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.

Reshetnikova LS, Garber MB. Crystallization of elongation factor G from an extreme thermophile Thermus thermophilus HB8. FEBS Lett. 1983; 154(2):149-150.

Karpova EA, Serdiuk IN, Tarkhovskiĭ IuS, Orlova EV, Boroviagin VL. Crystallization of ribosomes from Thermus thermophilus. Dokl Akad Nauk SSSR. 1986;289(5):1263-6.

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.

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.

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.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.

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.

Published

1987-05-20

Issue

Section

Short Communications