Albalat A, Mullen W, Husi H & Mischak H (2017) Tissue Proteomics in Vascular Disease. In: Touyz R & Schiffrin E (eds.) Hypertension: Tissue Proteomics in Vascular Disease. Methods in Molecular Biology, 1527. New York: Springer, pp. 53-60. http://link.springer.com/protocol/10.1007/978-1-4939-6625-7_4; https://doi.org/10.1007/978-1-4939-6625-7_4
In vascular diseases, as in many other pathophysiological conditions, tissue proteins are subjected to a number of different changes such as protein expression, posttranslational modifications, and proteolytic cleavage. For this reason, the study of the tissue proteome is becoming an increasingly important tool in biological and clinical research. In this chapter, we describe in detail the methodology for the analysis of tryptic digested peptides from aortic tissue extracts from mice with the aim to elucidate differences in the proteome between control and case or vascular diseased tissue samples. The method encompasses the analysis of these complex extracts in a single chromatographic run allowing for increased reproducibility. We also outline the main workflow for the processing of the generated data by statistical and bioinformatic analysis.
Tissue proteomics; FASP method; MS/MS data analysis