Tissue proteomics of the low-molecular-weight proteome using an integrated cLC-ESI-QTOFMS approach.
Proteomics. 2013 Mar 4;
Authors: Alvarez MT, Shah DJ, Thulin CD, Graves SW
Analysis of the protein/peptide composition of tissue has provided meaningful insights into tissue biology and even disease mechanisms. However, little has been published regarding top down methods to investigate lower molecular weight (MW) (500-5000 Da) species in tissue. Here we evaluate a tissue proteomics approach involving tissue homogenization followed by depletion of large proteins and then capillary liquid chromatography-mass spectrometry (cLC-MS) analysis to interrogate the low MW/low abundance tissue proteome. In the development of this method, sheep heart, lung, liver, kidney, and spleen were surveyed to test our ability to observe tissue differences. After categorical tissue differences were demonstrated, a detailed study of this method's reproducibility was undertaken to determine whether or not it is suitable for analyzing more subtle differences in the abundance of small proteins and peptides. Our results suggest that this method should be useful in exploring the low-MW proteome of tissues.
PMID: 23456981 [PubMed - as supplied by publisher]