A Top-Down/Bottom-Up Study of the Ribosomal Proteins of Caulobacter crescentus

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Date

2007

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Journal of Proteome Research

Abstract

Ribosomes from the Gram-negative α-proteobacterium Caulobacter crescentus were isolated using standard methods. Proteins were separated using a two-dimensional liquid chromatographic system that allowed the analysis of whole proteins by direct coupling to an ESI-QTOF mass spectrometer and of proteolytic digests by a number of mass spectrometric methods. The masses of 53 of 54 ribosomal proteins were directly measured. Protein identifications and proposed post-translational modifications were supported by proteolysis with trypsin, endoprotease Glu-C, and exoproteases carboxypeptidases Y and P. Tryptic peptide mass maps show an average sequence coverage of 62%, and carboxypeptidase C-terminal sequence tagging provided unambiguous identification of the small, highly basic proteins of the large subunit. C. crescentus presents some post-translational modifications that are similar to those of Escherichia coli (e.g., N-terminal acetylation of S9 and S18) along with some unique variations, such as a near absence of L7 and extensive modification of L11. The comprehensive description of this organism's ribosomal proteome provides a foundation for the study of ribosome structure, dependence of post-translational modifications on growth conditions, and the evolution of subcellular organelles.

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Keywords

ribosomal proteins, protein mass spectrometry, two-dimensional liquid chromatography, post-translational modifications

Citation

Running, W. E.; Ravipaty, S.; Karty, J. A.; Reilly, J. P. A Top-Down/Bottom-Up Study of the Ribosomal Proteins of Caulobacter Crescentus. J. Proteome Res. 2007, 6, 337-347.https://doi.org/10.1021/pr060306q

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Article