Mass Spectrometry Analysis of Proteome-wide Proteolytic Post-translational Degradation of Proteins

Journal Article · Fri Aug 01 04:00:00 EDT 2008 · Analytical Chemistry, 80(15):5819-5828

DOI:https://doi.org/10.1021/ac800077w· OSTI ID:947467

Shen, Yufeng; Hixson, Kim K.; Tolic, Nikola; Camp, David G.; Purvine, Samuel O.; Moore, Ronald J.; Smith, Richard D.

Protein proteolysis is an essential component to proper cell function. Here, we demonstrate a method for studying protein degradation by detection of intermediate intracellular peptides with a high-precision tandem mass spectrometry de novo sequencing-based approach. From a Saccharomyces cerevisiae lysate, we identified >1,200 peptides containing 6-100 amino acids without random false positives and ascribed most identifications as being products of protein degradation. Most protein degradation observed was located in the cytoplasm, and multiple types of cleavage were found to exist in addition to the expected trypsin-like and chymotrypsin-like preferences. The yeast nucleus was found as a proteolysis-inert organelle under the conditions studied and the V-ATPase to be degraded during disassembly. Additionally, matrix associated mitochondrial proteins functioning as transport carriers and gates were found to be commonly degraded. Determining these protein degradation events could eventually aid in understanding of cell biology and detection and treatment of protein degradation-related diseases.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 947467
• Report Number(s): PNNL-SA-58041; 26194; 20496; KP1704020
• Journal Information: Analytical Chemistry, 80(15):5819-5828, Journal Name: Analytical Chemistry, 80(15):5819-5828 Journal Issue: 15 Vol. 80; ISSN 0003-2700; ISSN ANCHAM
• Country of Publication: United States
• Language: English

Related Subjects

Environmental Molecular Sciences Laboratory
cellular biology
de novo sequencing
high precision ms/ms
protein degradation