Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR

Sang-Won Lee, Scott J. Berger, Suzana Martinović, Ljiljana Paša-Tolić, Gordon A. Anderson, Yufeng Shen, Rui Zhao, Richard D. Smith

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry coupled with capillary reverse-phase liquid chromatography was used to characterize intact proteins from the large subunit of the yeast ribosome. High mass measurement accuracy, achieved by "mass locking" with an internal standard from a dual electrospray ionization source, allowed identification of ribosomal proteins. Analyses of the intact proteins revealed information on cotranslational and posttranslational modifications of the ribosomal proteins that included loss of the initiating methionine, acetylation, methylation, and proteolytic maturation. High-resolution separations permitted differentiation of protein isoforms having high structural similarity as well as proteins from their modified forms, facilitating unequivocal assignments. The study identified 42 of the 43 core large ribosomal subunit proteins and 58 (of 64 possible) core large subunit protein isoforms having unique masses in a single analysis. These results demonstrate the basis for the high-throughput analyses of complex mixtures of intact proteins, which we believe will be an important complement to other approaches for defining protein modifications and their changes resulting from physiological processes or environmental perturbations.

Original languageEnglish
Pages (from-to)5942-5947
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume99
Issue number9
DOIs
Publication statusPublished - 2002 Apr 30

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Large Ribosome Subunits
Fungal Proteins
Ribosomal Proteins
Protein Isoforms
Proteins
Physiological Phenomena
Cyclotrons
Protein Subunits
Fourier Analysis
Reverse-Phase Chromatography
Post Translational Protein Processing
Acetylation
Complex Mixtures
Ribosomes
Methionine
Methylation
Mass Spectrometry
Yeasts
Ions

ASJC Scopus subject areas

  • Genetics
  • General

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Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR. / Lee, Sang-Won; Berger, Scott J.; Martinović, Suzana; Paša-Tolić, Ljiljana; Anderson, Gordon A.; Shen, Yufeng; Zhao, Rui; Smith, Richard D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, No. 9, 30.04.2002, p. 5942-5947.

Research output: Contribution to journalArticle

Lee, Sang-Won ; Berger, Scott J. ; Martinović, Suzana ; Paša-Tolić, Ljiljana ; Anderson, Gordon A. ; Shen, Yufeng ; Zhao, Rui ; Smith, Richard D. / Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR. In: Proceedings of the National Academy of Sciences of the United States of America. 2002 ; Vol. 99, No. 9. pp. 5942-5947.
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AU - Anderson, Gordon A.

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