Exploring the human seminal plasma proteome: an unexplored gold mine of biomarker for male infertility and male reproduction disorder

The human seminal fluid is a complex body fluid. It is not known how many proteins are expressed in the seminal plasma; however in analog with the blood it is possible up to 10,000 proteins are expressed in the seminal plasma. The human seminal fluid is a rich source of potential biomarkers for male infertility and reproduction disorder. In this review, the ongoing list of proteins identified from the human seminal fluid was collected. To date, 4188 redundant proteins of the seminal fluid are identified using different proteomics technology, including 2-DE, SDS-PAGE-LC-MS/MS, MudPIT. However, this was reduced to a database of 2168 non-redundant protein using UniProtKB/Swiss-Prot reviewed database. The core concept of proteome were analyzed including pT, MW, Amino Acids, Chromosome and PTM distribution in the human seminal plasma proteome. Additionally, the biological process, molecular function and KEGG pathway were investigated using DAVID software. Finally, the biomarker identified in different male reproductive system disorder was investigated using proteomics platforms so far. In this study, an attempt was made to update the human seminal plasma proteome database. Our finding showed that human seminal plasma studies used to date seem to have converged on a set of proteins that are repeatedly identified in many studies and that represent only a small fraction of the entire human seminal plasma proteome

Organelle isolation for proteomics: mitochondria from peripheral blood mononuclear cells

Mitochondria play key roles in many cell functions including energy production, fatty acid metabolism, pyrimidine biosynthesis, calcium homeostasis, and aging. They also regulate crucial signaling cascades such as apoptosis and oxidative stress. The proteome is often used to investigate the functional correlations on protein levels. Based upon the human, genome there is estimated 2000 to 2500 associated mitochondrial proteins, however, just over 600-800 have been identified at the protein level. For this reason, mitochondria contain a great number of proteins that have yet to be identified and characterized. The identification of these proteins can help in discovery of biological process. This protocol focuses on step-by-step procedure of mitochondrial proteome extraction from peripheral blood mononuclear cell [PBMC] mitochondria. The isolation and preparation procedures described here require 6 hours approximately