Two‐dimensional Fourier transform <sup>1</sup>H NMR studies of ribosomal protein E‐L30

Ven, F.J.M. van de; Bruin, Simon H. de; Hilbers, C. W.

doi:10.1016/0014-5793(84)80298-7

Cited by 15 publications

(11 citation statements)

References 9 publications

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“…16, and F. J. M. van de Ven, S. H. de Bruin, and C. W. Hilbers, personal communication) the secondary structural elements and their relative topology were deduced and are in excellent agreement with our findings.…”

supporting

confidence: 87%

Crystal structure of a prokaryotic ribosomal protein.

Wilson¹,

Appelt²,

Badger³

et al. 1986

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

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The structure of ribosomal protein L30 from Bacillus stearothermophilus has been solved to a resolution of 2.5 A. The molecule is somewhat elongated and contains two helices and a three-stranded, anti-parallel fl-pleated sheet. The protein fold, in which helices pack on the same side of the sheet, generates a simple helix-sheet, two-layered motif. It is possible to distinguish three hydrophobic patches on the molecular surface, and one end has six isolated arginine and lysine residues. It is proposed that these reflect sites of proteinprotein and protein-RNA interaction, respectively. The protein fold is very similar to that of the only other known ribosomal protein structure, L7/L12 from Escherichia coli, and, based on this similarity, an attempt is made to align the amino acid sequences of the two proteins.The ribosome is the cellular organelle, coordinating protein synthesis, that consists of a large and a small subunit. In prokaryotes, the intact ribosome is a 70S particle with a large SOS and a small 30S subunit. The 50S subunit has a 23S and a 5S RNA molecule and about 33 ("L") proteins; the 30S subunit has a 16S RNA molecule and about 20 ("S") proteins. We describe here the three-dimensional structure of protein L30 from Bacillus stearothermophilus. The ribosomal proteins from Escherichia coli have been the most extensively studied and are numbered according to their positions in two-dimensional gel electrophoresis (1). However, with the exception of L7/L12 (2), these proteins have failed to produce crystals suitable for x-ray diffraction studies, and we have concentrated on the proteins from the thermophile B. stearothermophilus, which are considerably more resistant to thermal denaturation (3). To avoid nomenclature confusion, homologies based on amino acid sequence data have been established between the available B. stearothermophilus proteins and those from E. coli, and the former are labeled accordingly. The L30 proteins have been fully sequenced (4,5) and are identical at 53% ofthe positions (Fig. 1). This degree of homology is typical of the ribosomal proteins from the two organisms and clearly shows that the considerable biochemical information that has been accumulated on the E. coli proteins can be directly applied to their counterparts from B. stearothermophilus.L30 is one of the smallest of the ribosomal proteins with a molecular weight of 7000. It is difficult to assign a particular biochemical function to L30 or, indeed, to any isolated ribosomal protein. It is not an essential protein since mutants that lack it have similar growth rates to the wild type (6). However, single protein omission experiments on reconstituted E. coli (7) and B. stearothermophilus (8, 9) ribosomes indicate a positive effect on peptidyltransferase activity. It appears, from reconstitution experiments (10), to be located near the surface of the large subunit and, from cross-linking experiments (11), to be close to the binding site of elongation factor Tu.Structure Determination. L30 was purified using a mild,...

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supporting

confidence: 87%

Crystal structure of a prokaryotic ribosomal protein.

Wilson¹,

Appelt²,

Badger³

et al. 1986

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

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“…The latter disappear to a large extent in 0.1 M NaCl. The same applies to resonances between 5 and 6 ppm, which arise from interactions between neighbouring strands of a &sheet [30]. (b) Protein C. There is a very large number of broadened amide resonances between 8 and 10 ppm, some perturbed aromatic resonances and an unusually large number of ring current shifted methyl resonances.…”

Section: Stokes Radius and Molecular Massmentioning

confidence: 93%

Purification and characterization of ribosomal proteins from the 30 S subunit of the extreme halophile Halobacterium marismortui

Shoham¹,

Dijk²,

Reinhardt³

et al. 1986

FEBS Letters

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Ribosomal proteins were extracted from 30 S subunits of Halobacterium marismortui under native conditions. Their separation was based on gel filtration and hydrophobic chromatography, performed at a concentration of 3.2 M KC1 to avoid denaturation. A total of nine proteins were isolated, purified and identified by partial amino-terminal sequences and two-dimensiona gel electrophoresis. There is a high degree of sequence homology with 30 S proteins from H. cutirubrum, and also some with 30 S proteins of eubacteria. Proton NMR data indicate unfolding of the proteins in low salt. One of the proteins, however, retains its secondary structure at a salt concentration as low as 0.1 M NaCl, and even in 8 M urea. One reason for this outstanding stability could be the high proportion (50%) of /?-structure in this protein as determined from circular dichroism measurements. In general, there is a higher B-sheet content than for 30 S proteins from Escherichia eoii. Measurements of Stokes radii indicate several of the proteins to have a rather elongated shape. One of these is a complex consisting of L3/L4 and L20, similar to the LI-complex from E. co&. The presence of this 50 S compiex in the preparation of the small subunit suggests a location on the interface between the subunits.(Halobacterium) Ribosomai protein 30 S subunit

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“…As is easily shown by density matrixs or product operator analysis,36 the amount of double quantum coherence produced by the first segment of sequence (24) is proportional to sin(2nJz), where J is the 13C-13C coupling constant. Thus, to observe one-bond couplings z is normally set to about 7ms, while for long-range couplings z may be several times this figure.…”

Section: D Inadequatementioning

confidence: 97%

Modern NMR techniques for structure elucidation

Morris

1986

Magnetic Reson in Chemistry

182

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The rapid growth during the last decade of multiple pulse methods in one (1D) and two-dimensional (2D) NMR has produced over 100 new techniques of potential use in high-resolution NMR. This review sets out to identify and to illustrate those new methods which are most useful in the determination of molecular structure. Ten basic techniques are considered, including 2D homo-and hetero-nuclear chemical shift correlation by direct and by relayed coherence transfer, and 1D polarization transfer. Numerous variations on the basic techniques, for example multiple quantum filtration, are also discussed. Particular attention is paid to the problems of sensitivity and of the optimum choice of experimental techniques for different classes of structural problem.

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Two‐dimensional Fourier transform ¹H NMR studies of ribosomal protein E‐L30

Cited by 15 publications

References 9 publications

Crystal structure of a prokaryotic ribosomal protein.

Crystal structure of a prokaryotic ribosomal protein.

Purification and characterization of ribosomal proteins from the 30 S subunit of the extreme halophile Halobacterium marismortui

Modern NMR techniques for structure elucidation

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Two‐dimensional Fourier transform 1H NMR studies of ribosomal protein E‐L30

Cited by 15 publications

References 9 publications

Crystal structure of a prokaryotic ribosomal protein.

Crystal structure of a prokaryotic ribosomal protein.

Purification and characterization of ribosomal proteins from the 30 S subunit of the extreme halophile Halobacterium marismortui

Modern NMR techniques for structure elucidation

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Product

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Two‐dimensional Fourier transform ¹H NMR studies of ribosomal protein E‐L30