Characterization of Protein Complexes using Targeted Proteomics

Author(s): Yassel Ramos Gomez, Sebastien Gallien, Vivian Huerta, Jan van Oostrum, Bruno Domon, Luis Javier Gonzalez.

Journal Name: Current Topics in Medicinal Chemistry

Volume 14 , Issue 3 , 2014

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Abstract:

Biological systems are not only controlled by the abundance of individual proteins, but also by the formation of complexes and the dynamics of protein-protein interactions. The identification of the components of protein complexes can be obtained by shotgun proteomics using affinity purification coupled to mass spectrometry. Such studies include the analyses of several samples and experimental controls in order to discriminate true specific interactions from unspecific interactions and contaminants. However, shotgun proteomics have limited quantification capabilities for low abundant proteins on large sample sets due to the undersampling and the stochastic precursor ion selection. In this context, targeted proteomics constitutes a powerful analytical tool to systematically detect and quantify peptides in multiple samples, for instance those obtained from affinity purification experiments. Hypothesis-driven strategies have mainly relied on the selected reaction monitoring (SRM) technique performed on triple quadrupole instruments, which enables highly selective and sensitive measurements of peptides, acting as surrogates of the pre-selected proteins, over a wide range of concentrations. More recently, novel quantitative methods based on high resolution instruments, such as the parallel reaction monitoring (PRM) technique implemented on the quadrupole-orbitrap instrument, have arisen and provided alternatives to perform quantitative analyses with enhanced selectivity.The application of targeted proteomics to protein-protein interaction experiments from plasma and other physiological fluid samples and the inclusion of parallel reaction monitoring (PRM), combined with other recent technology developments opens a vast area for clinical application of proteomics. It is anticipated that it will reveal valuable information about specific, individual, responses against drugs, exogenous proteins or pathogens.

Keywords: Affinity purification coupled with mass spectrometry, Protein-protein interactions, targeted proteomics, selected reaction monitoring, parallel reaction monitoring.

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Article Details

VOLUME: 14
ISSUE: 3
Year: 2014
Page: [344 - 350]
Pages: 7
DOI: 10.2174/1568026613666131204130124
Price: $58

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