Multiplexing Protein Analysis Core

CORE OVERVIEW

The Multiplexing Protein Analysis Laboratory uses LC-tandem MS with selected reaction monitoring (SRM) and LC-high resolution accurate mass selected ion monitoring (SIM) to measure protein abundance and isotope enrichment. The protein abundance measurements are unique because proteins are carefully selected and targeted for quantification. Targeting is achieved by designing methods to measure the abundance of several peptides. Adding deuterium oxide during the experimental period allows the determination of the synthesis rates of targeted proteins. An investigator using the core can then determine dynamic regulation of individual proteins.

Targeted Quantitative Proteomics

We use an LC-tandem mass spectrometry technique called selected reaction monitoring (SRM) to measure protein abundance. The assays are designed to target specific proteins based on their amino acid sequence, giving the assays high sensitivity in complex samples. Assays for individual proteins are bundled into panels of approximately 40 proteins that are designed to interrogate selected biochemical pathways.

Our workflow accommodates sufficient sample numbers to design experiments with good statistical power as well as a wide range of sample types and preparation methods. Users are asked to submit tissue or cell homogenates, made using tested methods from their lab, with precisely measured protein concentrations. We do the rest of the processing.

Synthesis and Turnover

Impaired protein homeostasis (proteostasis) is a hallmark of aging and is associated with several chronic diseases. Protein turnover, a primary proteostatic mechanism, is a dynamic process that consists of protein synthesis and protein degradation. To assess protein turnover as a proteostatic mechanism, the Core helps in the design and analysis of studies using deuterium oxide (D2O).

Core Leaders

Michael T. Kinter, Ph.D.

MPAC Lead

mike-kinter@omrf.org

Benjamin F. Miller, Ph.D.

MPAC Lead

benjamin-miller@omrf.org

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of Excellence in Aging Research

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