Amino acid sequence of a 50 S ribosomal protein involved in both EFG and EFT dependent GTP‐hydrolysis

Terhorst, Cox; Möller, W.; Laursen, Richard A.; Wittmann-Liebold, B.

doi:10.1016/0014-5793(72)80742-7

Cited by 73 publications

(28 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Two proteins from the small subunit, namely S5 and S18, do not yield N-terminal PTH amino acids when analyzed by the sequenator. They probably have blocked N-termini, as has already been found [6] for protein L7. From the 18 analyzed proteins of the small subunit, seven start with alanine, three with methionine, three with proline, two with serine and one each with either threonine, glycine or phenylalanine.…”

Section: Resultsmentioning

confidence: 54%

Studies on the primary structure of 20 proteins from Escherichia coli ribosomes by means of an improved protein sequenator

Wittmann‐Liebold¹

1973

FEBS Letters

View full text Add to dashboard Cite

Section: Resultsmentioning

confidence: 54%

Studies on the primary structure of 20 proteins from Escherichia coli ribosomes by means of an improved protein sequenator

Wittmann‐Liebold¹

1973

FEBS Letters

View full text Add to dashboard Cite

“…The value of uz was calculated rrom the amino acid composition sic to the fragment 1-73 [5]. Taking into account the distribution of hydrophobic amino acids in the N-terminal part of fragment 1 -73 and the fact that position 44 is occupied by proline [2], it may be assumed that the region responsible for dimerization is contained in the sequence 1-43.…”

Section: Discussionmentioning

confidence: 99%

“…The peculiarities of the protein L7/L12 primary structure [2] permit the isolation of different derivatives convenient for studying interactions between the subunits. Three methionine residues (14, 17 and 26) can be oxidized by hydrogen peroxide into their sulfoxides [3].…”

mentioning

confidence: 99%

The N‐Terminal Sequence Protein of L7/L12 Is Responsible for Its Dimerization

Gudkov¹,

Behlke

1978

European Journal of Biochemistry

View full text Add to dashboard Cite

Ultracentrifuge studies of intact protein L7/L12, of its fragments 27-120, 1-74 and 74-120 and of protein L7/L12 with oxidized methionine residues, indicate that the N-terminal sequence of the protein L7/L12 is responsible for its dimerization. The symmetry model of the dimer is discussed.Protein L7/L12 from Escherichia coli ribosomes is known to have a dimeric form in solution [l]. The peculiarities of the protein L7/L12 primary structure [2] permit the isolation of different derivatives convenient for studying interactions between the subunits. Three methionine residues (14, 17 and 26) can be oxidized by hydrogen peroxide into their sulfoxides [3]. Fragment 27-120 can be obtained by treatment with cyanogen bromide. The presence of the only arginine residue in position 73 allows the isolation of the fragments 1-73 and 74-120.Ultracentrifugation studies of the protein L7/L12 and its fragments led to the conclusion that the region responsible for its dimerization is located in its Nterminal sequence. MATERIALS AND METHODS The buffer solutions: buffer Isolation and Purification of Protein L7/L12The fraction of acidic ribosomal proteins was isolated by column chromatography with carboxymethylcellulose [4]. It was desalted on a Sephadex G-25 column in buffer D, lyophilized and further separation was carried out in conditions as described [l]. To remove aggregates, the proteins L7/L12 were subjected to column chromatography with Sephadex G-100 in buffer D. Oxidation of Protein L7/L12In order to oxidize methionine residues the protein was dissolved in a small amount of buffer D ; 50 % acetic acid containing hydrogen peroxide was added and the mixture was kept overnight at 4 "C. The final concentration of protein in solution was 2-3 mg/ml and that of hydrogen peroxide 0.7 -1 .O mg/ml.

show abstract

“…Moreover, rp S3 was found to act as an endonuclease in the DNA-repairing system [2]. Finally, some data indicate that tumor diseases may be accompanied by overexpression of rp genes 13,4) and by chromosomal rearrangements in regions of rp genes [ S ] . Therefore, structural analysis of these proteins is necessary for complete understanding of their fundamental role in the cell.…”

mentioning

confidence: 99%

Characterization of the Human Small‐Ribosomal‐Subunit Proteins by N‐Terminal and Internal Sequencing, and Mass Spectrometry

Vladimirov

Ivanov

Карпова

et al. 1996

European Journal of Biochemistry

View full text Add to dashboard Cite

Reverse-phase HPLC was used to fractionate 40s ribosomal proteins from human placenta. Application of a C, reverse-phase column allowed us to obtain 27 well-resolved peaks. The protein composition of each chromatographic fraction was established by two-dimensional polyacrylamide gel electrophoresis and N-terminal sequencing. N-terminally blocked proteins were cleaved with endoproteinase Lys-C, and suitable peptides were sequenced. All sequences were compared with those of ribosomal proteins available from data bases. This allowed us to identify all proteins from the 40s human ribosomal subunit in the HPLC elution profile. By matrix-assisted laser-desorption ionization mass spectrometry the masses of the 40s proteins were determined and checked for the presence of post-translational modifications. For several proteins differences to the deduced sequences and the calculated masses were found to be due to post-translational modifications.Keywords: human ribosomal proteins ; HPLC ; N-terminal and internal amino acid sequencing ; matrixassisted-laser-desorption ionization MS ; post-translational modifications.Human ribosornd proteins are the main components of the fundamental processes which translate genetic information into proteins at the ribosome. Some ribosomal proteins (rp) are further involved in non-ribosomal functions. Recently, rp L7 was shown to contain the basic-region-leucine-zipper sequence which mediates the stable binding of this protein to DNA [I]. Moreover, rp S3 was found to act as an endonuclease in the DNA-repairing system [2]. Finally, some data indicate that tumor diseases may be accompanied by overexpression of rp genes 13, 4) and by chromosomal rearrangements in regions of rp genes [ S ] . Therefore, structural analysis of these proteins is necessary for complete understanding of their fundamental role in the cell. The task of structure determination of rat rp has been almost fully resolved by the application of recombinant-DNA technology ; details are reviewed in [6]. However, knowledge of the cDNA sequences does not provide information concerning post-translational modifications of the proteins after their biosynthesis and assembly into the ribosomal subunits. Several modifications of rp have been determined in the eubacterial ribosome [7, 81, and the eukaryotic rprotein S6 is known to be phosphorylated 191. This phosphorylation was shown to be connected with cell growth [I 01 and cell cycle control [I I]. At least seven 40s proteins from HeLa cells were found to be methylated, and the level of their methylation was shown to be altered in the Abbreviations. MALDI, matrix-assisted laser-desorption ionization ; Lys-C, endoproteinase Lys-C; RP, reverse-phase; rp, ribosomal protein ; S-proteins, proteins of the small ribosomal subunit; 40S, small ribosomal subunit; TP40, total protein mixture of the small ribosomal subunit; TOF, time-of-flight ; Pth-Xaa, phenylthiohydantoin -amino-acid.course of the cell cycle [12]. It is also known that ribosomal proteins may be acetylated 17, 13, 141. However...

show abstract

Amino acid sequence of a 50 S ribosomal protein involved in both EFG and EFT dependent GTP‐hydrolysis

Cited by 73 publications

References 23 publications

Studies on the primary structure of 20 proteins from Escherichia coli ribosomes by means of an improved protein sequenator

Studies on the primary structure of 20 proteins from Escherichia coli ribosomes by means of an improved protein sequenator

The N‐Terminal Sequence Protein of L7/L12 Is Responsible for Its Dimerization

Characterization of the Human Small‐Ribosomal‐Subunit Proteins by N‐Terminal and Internal Sequencing, and Mass Spectrometry

Contact Info

Product

Resources

About