title: Protein expression profiles of human lymph and plasma mapped by 2D-DIGE and 1D SDS-PAGE coupled with nanoLC-ESI-MS/MS bottom-up proteomics.
abstract: In this study a proteomic approach was used to define the protein content of matched samples of afferent prenodal lymph and plasma derived from healthy volunteers. The analysis was performed using two analytical methodologies coupled with nanoliquid chromatography-tandem mass spectrometry: one-dimensional gel electrophoresis (1DEF nanoLC Orbitrap-ESI-MS/MS), and two-dimensional fluorescence difference-in-gel electrophoresis (2D-DIGE nanoLC-ESI-MS/MS). The 253 significantly identified proteins (p<0.05), obtained from the tandem mass spectrometry data, were further analyzed with pathway analysis (IPA) to define the functional signature of prenodal lymph and matched plasma. The 1DEF coupled with nanoLC-MS-MS revealed that the common proteome between the two biological fluids (144 out of 253 proteins) was dominated by complement activation and blood coagulation components, transporters and protease inhibitors. The enriched proteome of human lymph (72 proteins) consisted of products derived from the extracellular matrix, apoptosis and cellular catabolism. In contrast, the enriched proteome of human plasma (37 proteins) consisted of soluble molecules of the coagulation system and cell-cell signaling factors. The functional networks associated with both common and source-distinctive proteomes highlight the principal biological activity of these immunologically relevant body fluids.
authors.affiliation: Department of Pathology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Cristina C
authors.initials: CC
authors.lastname: Clement
authors.forename: David
authors.initials: D
authors.lastname: Aphkhazava
authors.forename: Edward
authors.initials: E
authors.lastname: Nieves
authors.forename: Myrasol
authors.initials: M
authors.lastname: Callaway
authors.forename: Waldemar
authors.initials: W
authors.lastname: Olszewski
authors.forename: Olaf
authors.initials: O
authors.lastname: Rotzschke
authors.forename: Laura
authors.initials: L
authors.lastname: Santambrogio
mesh_terms: D000328:Adult; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008196:Lymph / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D010949:Plasma / Q000378:metabolism*; D020543:Proteome / Q000096:biosynthesis*; D040901:Proteomics / Q000379:methods*
chemical_list: D020543:Proteome