Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR
- 30 April 2002
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 99 (9) , 5942-5947
- https://doi.org/10.1073/pnas.082119899
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.Keywords
This publication has 51 references indexed in Scilit:
- Matrix-assisted ultraviolet laser desorption of non-volatile compoundsPublished by Elsevier ,2001
- The Structural Basis of Ribosome Activity in Peptide Bond SynthesisScience, 2000
- A comparison of the yeast and rabbit 80 S ribosome reveals the topology of the nascent chain exit tunnel, inter-subunit bridges and mammalian rRNA expansion segmentsJournal of Molecular Biology, 2000
- Routine Part-per-Million Mass Accuracy for High- Mass Ions: Space-Charge Effects in MALDI FT-ICRAnalytical Chemistry, 1998
- How the Ribosome WorksAmerican Scientist, 1998
- Mass Spectrometric Analysis of 40 S Ribosomal Proteins from Rat-1 FibroblastsPublished by Elsevier ,1996
- Partial purification and characterization of the specific protein-lysine N-methyltransferase of YL32, a yeast ribosomal proteinBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1989
- Two‐dimensional 1H‐NMR investigation of ribonuclease T1European Journal of Biochemistry, 1988
- Space charge effects in Fourier transform mass spectrometry. II. Mass calibrationAnalytical Chemistry, 1984
- OCCURRENCE, ISOLATION, AND CHARACTERIZATION OF POLYRIBOSOMES IN YEASTThe Journal of cell biology, 1967