Protein involved in the binding of dihydrostreptomycin to ribosomes of Escherichia coli

Schreiner, George F.; Nierhaus, K H

doi:10.1016/0022-2836(73)90248-9

Cited by 95 publications

(46 citation statements)

References 30 publications

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“…It has been reported that streptomycin binding requires the presence of protein S12 and that the 320 drug binds to a site formed by proteins S3 and S5 [17] or directly to 165 rRNA [19]. One possible explanation of our binding data could be that neither S12, S4, SS are constituent proteins of the binding site but that the latter may be shielded by alterations in protein S12 [ 191 and reformed by altered S4 or S5 proteins.…”

Section: Resultsmentioning

confidence: 77%

Ribosomal ambiguity (ram) mutations facilitate dihydrostreptomycin binding to ribosomes

Böck

Piepersberg

1979

FEBS Letters

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Section: Resultsmentioning

confidence: 77%

Ribosomal ambiguity (ram) mutations facilitate dihydrostreptomycin binding to ribosomes

Böck

Piepersberg

1979

FEBS Letters

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“…This group contains the 3 0 4 proteins S1, S2, S3, S5, S9, S10, S14, and S21 [9,13]. Only three (S1, S2, and Sl0) of these eight proteins were found in the S-150 fractions (Table l), whereas S6, another "pool protein", belongs to the proteins firmly bound to the ribosome [9]. [12,19] are too controversial to be used for making correlation with pool size.…”

Section: Proteinmentioning

confidence: 99%

“…1 7; Bacto tryptone (Difco), 0.5% yeast extract (Difco); 0.09 M NaCl, 0.001 N NaOH and 0.2% glucose, and harvested at about 5 x lo8 cells per ml. Ribosomes were prepared as described [9]. After pelleting the ribosomes the supernatant was extensively dialyzed against a buffer containing 10 mM Tris .…”

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confidence: 99%

Pools of Ribosomal Proteins in Escherichia coli. Studies on the Exchange of Proteins between Pools and Ribosomes

Ulbrich

Nierhaus

1975

Eur J Biochem

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A new labelling technique in vivo has been introduced which allows the tritiation of cell components with high specific activity during growth in rich medium. By this technique the pool size of each protein can be measured directly in the supernatant from centrifugation at 150 000 x g. A measurable pool was found for the proteins S1, S2, S6, S10, L1, L4, L7, L8/9, L10, L12, L21, and L25.Experiments on migration of ribosomal proteins from the supernatant to ribosomes (i.e. association) and vice versa (dissociation) demonstrate a remarkable constancy in the composition of the ribosome. There is no significant difference between ribosomes engaged or not engaged in poly-(Phe) synthesis.Attempts to measure the pool size of ribosomal proteins can be divided into two classes. (a) In cells grown in rich medium the pools of ribosomal proteins were analyzed by immunological techniques [I] ; a remarkably high fraction (7 to 9 o/,) of total ribosomal proteins was found in the pools. (b) In cells labelled with radioactive amino acids the pools of individual ribosomal proteins were calculated [2 -61 ; to guarantee a reasonable uptake of label in these experiments, the cells had to be grown in a poor medium; a low pool size for the ribosomal proteins (less than 4%) were found in these studies. The discrepancy between the two classes of experiments can, at least partially, be explained by the different growth rates of cells in rich and poor media. The pool size of ribosomal proteins increases with decreasing generation time [l, 61. The same relation between pool size and growth rate was reported to be valid for pools of ribosomal precursors [7].In this paper we describe a technique which enables us to measure the individual pool size for each ribosomal protein during exponential growth in rich medium. Furthermore, exchange of ribosomal proteins between ribosomes and pools was measured under various conditions. It was found that the protein exchange between pools and ribosomes did not Abhreviatiunb. Poly(U), poly(uridylic acid) ; poly(Phe), polyphenylalanine.differ significantly in the presence or absence of poly(Phe) synthesis. MATERIALS AND METHODS Preparation of Ribosomes and S-150 FractionCells from Escherichiu coZi K12 (strain A19) were grown in L medium [8], i.e. 1 7; Bacto tryptone (Difco), 0.5% yeast extract (Difco); 0.09 M NaCl, 0.001 N NaOH and 0.2% glucose, and harvested at about 5 x lo8 cells per ml. Ribosomes were prepared as described [9]. After pelleting the ribosomes the supernatant was extensively dialyzed against a buffer containing 10 mM Tris . HCl, pH 7.5, 10 mM MgC1, and 6 mM 2-mercaptoethanol and centrifuged again at 176000 x g for 3 h. The supernatant was taken as the S-150 fraction. In the poly(U) system (see next section) the S-150 fraction showed background activity; this activity was stimulated more than 200-fold by adding ribosomes.For labelling the cells, tritiated Bacto tryptone (Difco; tritiated by Amersham with the Wilzbach method; 2.5 mCi/g) and yeast extract (Difco ; tritiated by Amcrsham with t...

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“…These include chemical crosslinking of neighboring proteins (e.g., refs. 7 and 8), chemical protection (9), fluorescent energy transfer (10,11), affinity labeling (12)(13)(14)(15)(16)(17), and neutron scattering (18 We have built such a model by using the following two-step procedure. First, a three-dimensional plaster model was made based on the immuno-EM diagrams of Tischendorf et al (1,2) and the antigenic sites were marked on its surface.…”

mentioning

confidence: 99%

Use of computerized multidimensional scaling to compare immunoelectron microscopy data with protein near-neighbor information: application to the 30S ribosome from Escherichia coli.

Gaffney¹,

Craven²

1978

Proc. Natl. Acad. Sci. U.S.A.

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A three-dimensional model of the protein arrangement in the Escherichia coli 30S ribosome was constructed by using computerized multidimensional scaling of immunoelectron microscope data. This enabled data comparison between the new electron microscope technique and other methods such as crosslinking, chemical protection, affinity labeling, energy transfer, and assembly interactions. The immunoelectron microscopy data are reasonably consistent with those from other sources. Reasons for some inconsistent data are discussed and our calculation of the dimensions of the proteins, both globular and elongated, are summarized. One of the most interesting recent developments in the analysis of ribosome structure is the use of immunoelectron microscopy (immuno-EM) to determine the relative positions of the individual proteins on the surface of the subunit particle (1)(2)(3)(4)(5) We have built such a model by using the following two-step procedure. First, a three-dimensional plaster model was made based on the immuno-EM diagrams of Tischendorf et al. (1, 2) and the antigenic sites were marked on its surface. Second, the shortest distances between all pairs of these antigenic sites were measured and used as input for a multidimensional scaling program. Its output gives the three-dimensional coordinates for a "naked" protein model-one without the outline of the ribosome. The model provides a unique and insightful view of the 30S ribosome. The uneven distribution of proteins within the subunit and the elongation of 12 of them are strikingly apparent. Questions on the relationship of structure to function take on fresh meaning but, more importantly, the model allows us to make a systematic comparison of all the relevant published data regarding protein-protein relationships.Computer program Multidimensional scaling is a statistical technique that has been used for approximately 15 years in psychology and is now being used extensively in other fields, including an early attempt to generate a three-dimensional model of the 30S ribosome (19). It constructs a configuration of points in space from information about the distances between points. For example, if a matrix were made of the 703 distances in miles between any 38 cities in the United States, and used as input in a multidimensional scaling program, the output would be 38 points representing the cities in the correct two-dimensional configuration. The orientation (north, south, east, west) would be left for the subjective decision of the user. Missing data and tied data can be accommodated but present some problems.The program places N points in a space of a given dimension so as to minimize STRESS, which measures the badness-of-fit between the configuration of points and the data. It represents the extent to which the data deviate in the least squares sense from the monotonic curve (see Fig. 3 A and B)

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Protein involved in the binding of dihydrostreptomycin to ribosomes of Escherichia coli

Cited by 95 publications

References 30 publications

Ribosomal ambiguity (ram) mutations facilitate dihydrostreptomycin binding to ribosomes

Ribosomal ambiguity (ram) mutations facilitate dihydrostreptomycin binding to ribosomes

Pools of Ribosomal Proteins in Escherichia coli. Studies on the Exchange of Proteins between Pools and Ribosomes

Use of computerized multidimensional scaling to compare immunoelectron microscopy data with protein near-neighbor information: application to the 30S ribosome from Escherichia coli.

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