Direct Mass Spectrometric Analysis of Intact Proteins of the Yeast Large Ribosomal Subunit using Capillary LC/FTICR
- BATTELLE (PACIFIC NW LAB)
- ASSOC WESTERN UNIVERSITY
Electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry coupled with capillary reverse phase LC (RPLC) 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 ESI source, allowed identification of ribosomal proteins. Analyses of the intact proteins revealed information on co-translational and post-translational 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.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- US Department of Energy (US)
- DOE Contract Number:
- AC06-76RL01830
- OSTI ID:
- 15001290
- Report Number(s):
- PNWD-SA-5450; TRN: US200404%%66
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, Issue 9; Other Information: PBD: 30 Apr 2002
- Country of Publication:
- United States
- Language:
- English
Similar Records
Quantitative Analysis of Human Salivary Gland-Derived Intact Proteome Using Top-Down Mass Spectrometry
High Speed Intact Protein Characterization Using 4X Frequency Multiplication, Ion Trap Harmonization, and 21 Tesla FTICR-MS