Characterization and Analysis of Posttranslational Modifications of the Human Large Cytoplasmic Ribosomal Subunit Proteins by Mass Spectrometry and Edman Sequencing

  title={Characterization and Analysis of Posttranslational Modifications of the Human Large Cytoplasmic Ribosomal Subunit Proteins by Mass Spectrometry and Edman Sequencing},
  author={Tatyana I. Odintsova and E C M{\"u}ller and Anton V. Ivanov and Tsezi A. Egorov and Ralf Bienert and Serguei Vladimirov and Susanne Kostka and Albrecht Otto and Brigitte Dr Wittmann-Liebold and Galina G. Karpova},
  journal={Journal of Protein Chemistry},
The 60S ribosomal proteins were isolated from ribosomes of human placenta and separated by reversed phase HPLC. The fractions obtained were subjected to trypsin and Glu-C digestion and analyzed by mass fingerprinting (MALDI-TOF), MS/MS (ESI), and Edman sequencing. Forty-six large subunit proteins were found, 22 of which showed masses in accordance with the SwissProt database (June 2002) masses (proteins L6, L7, L9, L13, L15, L17, L18, L21, L22, L24, L26, L27, L30, L32, L34, L35, L36, L37, L37A… 

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Mutations Preventing the Phosphorylation of Human Ribosomal Protein uS15 at Y38 and S48 Reduce the Efficiency of its Transfer into the Nucleolus

The findings suggest the importance of phosphorylation of the ribosomal protein uS15 by cytoplasmic protein kinases at several sites for its efficient transfer into the nucleolus, where pre-ribosomal subunits are assembled.

Replacement of Hydroxylated His39 in Ribosomal Protein uL15 with Ala or Thr Impairs the Translational Activity of Human Ribosomes

It has been found that exogenous uL 15 3xFLAG is able to functionally replace endogenous uL15 in HEK293 cells transfected with an appropriate DNA construct and this stabilization of the ribosome structure ensures the maintenance of its translational activity.

Analysis of the Arabidopsis Cytosolic Ribosome Proteome Provides Detailed Insights into Its Components and Their Post-translational Modification *S

In silico digestion of all 409 ribosomal protein sequences in Arabidopsis defined the proportion of theoretical gene-specific peptides for each gene family and highlighted the need for low m/z cutoffs of MS ion selection for MS/MS to characterize low molecular weight, highly basic ribosome proteins.

Human METTL18 is a histidine-specific methyltransferase that targets RPL3 and affects ribosome biogenesis and function

METTL18 is established as the second human histidine-specific protein MTase, and its functional relevance is demonstrated, indicating that METTL18-mediated methylation of RPL3 is important for optimal ribosome biogenesis and function.

Identification and Biological Characterization of Ribosomal Protein Methyltransferases in Yeast and Humans

This dissertation aimed to identify and characterize ribosomal protein methyltransferases in S. cerevisiae and humans and uncover their biological roles, finding that Hpm1 is a multifunctional enzyme with independent roles in ribosome biogenesis and translation; the latter regulated by Rpl3 methylation and the former by methylation of yet unknown proteins.

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Mass Spectrometric Analysis of 40 S Ribosomal Proteins from Rat-1 Fibroblasts*

This study establishes a powerful method to rapidly analyze protein components of large biological complexes and their covalent modifications.

Post-translational processing of rat ribosomal proteins. Ubiquitous methylation of Lys22 within the zinc-finger motif of RL40 (carboxy-terminal extension protein 52) and tissue-specific methylation of Lys4 in RL29.

The rat RL40 structure was identical to the cDNA-predicted sequence except for complete stoichiometric N epsilon-trimethylation of Lys22 within its zinc-finger motif; this modification occurred in the ribosomes of all three rat tissues investigated but not in yeast ribosome.

Characterization of the human small-ribosomal-subunit proteins by N-terminal and internal sequencing, and mass spectrometry.

Reverse-phase HPLC was used to fractionate 40S ribosomal proteins from human placenta and found differences to the deduced sequences and the calculated masses were found to be due to post-translational modifications.

Determination of the complete amino-acid sequence of protein S6 from the wild-type and a mutant of Escherichia coli.

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Identification and Sequence Analysis of Contact Sites between Ribosomal Proteins and rRNA in Escherichia coli 30 S Subunits by a New Approach Using Matrix-assisted Laser Desorption/Ionization-Mass Spectrometry Combined with N-terminal Microsequencing*

The novel approach applied here promises to be useful for similar studies on other known protein·RNA complexes and for the first time the cross-linked rRNA moiety to be sequenced by this technique.

Observation of Escherichia coli ribosomal proteins and their posttranslational modifications by mass spectrometry.

Mass spectral peak locations were consistent with previously reported post-translational modifications involving N-terminal methionine loss, methylation, thiomethylation, and acetylation for all but one case.

The Action of N-terminal Acetyltransferases on Yeast Ribosomal Proteins*

It is suggested that the acetylation of NatD substrates requires not only Ard1p andNat1p, but also auxiliary factors that are acetylated by the Mak3p and Nat3p N-terminal acetyltransferases.

Methylation of ribosomal proteins during ribosome assembly in Escherichia coli

  • F. Chang
  • Biology
    Molecular and General Genetics MGG
  • 2004
The extent of methylation of both L7 and L11 stays nearly constant during the cell growth cycle from early log to stationary phase and the methyl groups of L11 appear to be stable at either 25°C or 37°C.

High-throughput mass spectrometric discovery of protein post-translational modifications.

The approach is amenable to automation, it presents a potentially large-scale means of protein characterisation in proteome projects, and is illustrated with proteins from 2-D gels of Escherichia coli and sheep wool, where post-translational modifications predicted by FindMod were confirmed by MALDI post-source decay peptide fragmentation.