Age related post translational modifications to proteins that alter protein complexes determined by FTICR-MS
EMSL Project ID
12291a
Abstract
Reactive oxygen species (ROS) are central to the immune response; however, ROS also produce posttranslational modifications (PTMs) on intracellular proteins in the state of chronic inflammation we call aging. Some age-related pathologies are associated with PTMs to proteins, one such protein known to be oxidized with aging is the calcium regulatory protein calmodulin (CaM). We have shown that oxidative stress in cells induces repair processes which remove PTMs from CaM and we found an oxidation dependent C-terminal lysine cleavage to CaM which can be used as a stable biomarker of intracellular oxidative stress. Thus an in depth characterization of the changes in PTMs to intercellular proteins with oxidative stress (e.g. inflammation or aging) and the discovery of how these modifications alter protein complexes, will further our understanding of how PTMs contribute to the aging and disease processes. The specific experimental goals of this proposal are to determine the age dependent changes in oxidatively induced PTMs to proteins; and resolve PTMs induced alterations in protein-protein interactions using high resolution separations and high performance mass spectrometry. Understanding the changes that occur in the brain proteome with aging will be instrumental in defining the underlying cause of some of age related pathologies and related inflammatory disease. We propose to identify age related changes in the proteome and PTMs within it utilizing a high throughput global approach eliminating time consuming low resolution methodologies.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2007-06-05
End Date
2009-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Related Publications
Lopez-Ferrer D, KK Hixson, HS Smallwood, TC Squier, K Petritis, and RD Smith. 2009. "Evaluation of a High Intensity Focused Ultrasound-Immobilized Trypsin Digestion and 18 O-Labeling Method for Quantitative Proteomics." Analytical Chemistry 81(15):6272-6277.
Smallwood HS, D Lopez-Ferrer, and TC Squier. 2011. "Aging Enhances the Production of Reactive Oxygen Species and Bactericidal Activity in Peritoneal Macrophages by Upregulating Classical Activation Pathways." Biochemistry 50(45):9911-9922. doi:10.1021/bi2011866