The crystallization of protein BL17 from the 50 S ribosomal subunit of <i>Bacillus stearothermophilus</i>

Appelt, K.; Dijk, Jan; Epp, Otto

doi:10.1016/0014-5793(79)81251-x

Cited by 26 publications

(10 citation statements)

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“…replacement (MIR) method (Hoffman et al, 1994). The B. stearothermophilus protein was chosen for study because of its high degree of sequence homology to the E. coli protein, and its amenability to crystallization (Appelt et al, 1979). In our earlier crystal structure, L9 was found to have a highly elongated and unusual structure consisting of two domains joined by a long a-helix.…”

Section: Introductionmentioning

confidence: 99%

Ribosomal Protein L9: A Structure Determination by the Combined Use of X-ray Crystallography and NMR Spectroscopy

Hoffman

Cameron

Davies

et al. 1996

Journal of Molecular Biology

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Section: Introductionmentioning

confidence: 99%

Ribosomal Protein L9: A Structure Determination by the Combined Use of X-ray Crystallography and NMR Spectroscopy

Hoffman

Cameron

Davies

et al. 1996

Journal of Molecular Biology

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“…6 Thanks to work at Weizmann and Max Plank the first micro crystals were obtained in the early 1980s. 7 Professor Yonath was inspired by an article she read at the time about how the cells of winter sleeping bears pack ribosomes in an orderly way and concluded that under stressful conditions this is a natural mechanism for preserving non-deteriorated ribosomes. Therefore she decided to use ribosomes from organisms living in harsh conditions.…”

Section: Ribosome Studiesmentioning

confidence: 99%

Preface: Science and Public Health:Professor Ada Yonath, Weizmann Institute of Science, Rehovot, Israel, Nobel Laureate in Chemistry, 2009

Leighton

2012

Public Health Rev

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Ribosomes are responsible for translating genetic code into protein and thus central components of cell life. Knowledge of their structure provided important potential applications in and understanding of the mode of action of antibiotics and resistance to antibiotics. The work of Professor Ada Yonath on the crystallography of ribosomes led to improvements in crystallization, crystallographic techniques, and x-ray diffraction detection procedures. In 2000 and 2001, Dr. Yonath published the three dimensional structures of the two subunits of bacterial ribosome. There are potential applications for this work, and hopefully science will use such tools to develop new methods to reduce the burden of many diseases. In 2009, Professor Ada Yonath was awarded the Nobel Prize in Chemistry for her structural work on the ribosome.

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“…However, one cannot conclude from these observations that BL17 is a non-ribosomal protein since washing of E. coli ribosomes with sucrose and salt reduces the intensity of many ribosomal proteins [ 14,151 and since ribosomes from E. coli mutants missing one (or even more) protein(s) are nevertheless active in protein biosynthesis [ 16-191. Furthermore, the BL17 preparation used in these experiments was isolated from 70 S ribosomes which had been washed with sucrose and salt and was obtained in amounts comparable to those of the other ribosomal, proteins. The weak BL17 spot on two-dimensional gels could be attributed to variable binding of the dye [7].…”

Section: Comparison With Other Ribosomal Proteinsmentioning

confidence: 99%

“…Noteworthy features of BL17 are that tryptophan and cysteine are absent, a lysine-rich region is present in the N-terminal part @OS. [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19], and the hydrophobic amino acids are not clustered but are evenly distributed along the protein chain.…”

Section: Characteristics Of the Sequencementioning

confidence: 99%

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The primary structure of protein BL17 isolated from the large subunit of the Bacillus stearothermophilus ribosome

1980

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To understand, at the molecular level, the function of the ribosome which plays a central role in the biosynthesis of proteins, it is essential to know the structure of the particle in great detail. So far the primary structures of almost all of the 53 proteins [I] and the 3 RNA molecules [Z] present in the Escherichiu coli ribosome are known. Progress has been made towards the elucidation of secondary structures of both the proteins [l] and the RNAs [3-51 also.The most direct way of determining the tertiary structure of a protein is by its crystallization followed by X-ray analysis. This approach can now be applied to ribosomal proteins. The C-terminal fragment of E. coli ribosomal proteins L7/L12 has been crystallized [6] and analyzed at a resolution of 2.6 A (A. Liljas, persond commu~cation). The first intact ribosomal protein which could be crystallized f7] is protein BL17 isolated from the large subunit of the Bacillus, sfeurothermophilus ribosome. The determination of the tertiary structure of this protein by X-ray analysis is now in progress (0. Epp and R. Reinhardt, unpubllshed) .Here, we describe the complete primary structure of protein BL17 with 147 amino acids and present its secondary structure based on 4 prediction programmes. The ammo acid sequence of BL17 is compared to that of other ribosomal proteins. Materials and methodsProtein BL17 was isolated as in [S] avoiding urea and acetic acid extraction. Enzymic digestion was done as follows: ElsevierfNorth-Holland Biomedical Press(i) Trypsin digestion at pH 8 .l (in 0.2 M N-methylmorpholine acetate buffer at 37OC for 3 h); (ii) ArmiZZaria mellea protease digestion at pH 8.1(same buffer as above at 37°C for 6 h); (iii) Pepsin digestion at pH 2 (in 5% formic acid at 37'C for 6 h); (iv) S~a~hyZococcus aureus protease digestion at pH 4.0 (in 0.1 M ammonium acetate buffer at 37°C for 48 h). The resulting peptides were isolated by: (i) One-dimensional preparative thin-layer chromatography (A. metlea protease peptides); (ii) Thin-layer fingerprints (tryptic peptides); (iii) Gel-filtration on Sephadex GSO, superfine(1 X 140 cm) in 10% acetic acid followed by fmgerprints on thin-layer plates (peptic and Staph. aureus peptides). Sequencing of the various peptides was done with the DABITC~~C double-coupling method 19 3. DABITH-Ile and -Leu were determined by onedimensional thin-layer chromatography on polyamide sheets [lo]. Amino acid analysis was done on Durrum D-500 analysersasin [ll]. Results and discussion .l . Sequence determinationTwenty-four peptides were isolated from the tryptic digestion of BLI 7 by thin-layer frn~~~nt~g, and the complete sequences of these peptides (except T7, TlO and T15) were determined. The sequences of the peptides T7 (pas. 32-44), and the order of the tryptic pep tides were obtained from the sequences of the peptic peptides. Sixteen peptides (PI-P16) were isolated 323

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The crystallization of protein BL17 from the 50 S ribosomal subunit of Bacillus stearothermophilus

Cited by 26 publications

References 12 publications

Ribosomal Protein L9: A Structure Determination by the Combined Use of X-ray Crystallography and NMR Spectroscopy

Ribosomal Protein L9: A Structure Determination by the Combined Use of X-ray Crystallography and NMR Spectroscopy

Preface: Science and Public Health:Professor Ada Yonath, Weizmann Institute of Science, Rehovot, Israel, Nobel Laureate in Chemistry, 2009

The primary structure of protein BL17 isolated from the large subunit of the Bacillus stearothermophilus ribosome

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