title: Longitudinal Urinary Protein Variability in Participants of the Space Flight Simulation Program.
abstract: Urine is a valuable material for the diagnosis of renal pathologies and to investigate the effects of their treatment. However, the variability in protein abundance in the context of normal homeostasis remains a major challenge in urinary proteomics. In this study, the analysis of urine samples collected from healthy individuals, rigorously selected to take part in the MARS-500 spaceflight simulation program, provided a unique opportunity to estimate normal concentration ranges for an extended set of urinary proteins. In order to systematically identify and reliably quantify peptides/proteins across a large sample cohort, a targeted mass spectrometry method was developed. The performance of parallel reaction monitoring (PRM) analyses was improved by implementing tight control of the monitoring windows during LC-MS/MS runs, using an on-the-fly correction routine. Matching the experimentally obtained MS/MS spectra with reference fragmentation patterns allowed dependable peptide identifications to be made. Following optimization and evaluation, the targeted method was applied to investigate protein abundance variability in 56 urine samples, collected from six volunteers participating in the MARS-500 program. The intrapersonal protein concentration ranges were determined for each individual and showed unexpectedly high abundance variation, with an average difference of 1 order of magnitude.
authors.affiliation: Luxembourg Clinical Proteomics Center (LCP), Luxembourg Institute of Health , Strassen 1445, Luxembourg.|University of Luxembourg , Esch-sur-Alzette 4365, Luxembourg.
authors.forename: Nina A
authors.initials: NA
authors.lastname: Khristenko
authors.affiliation: Institute for Biomedical Problems , Moscow 123007, Russia.
authors.forename: Irina M
authors.initials: IM
authors.lastname: Larina
authors.affiliation: Luxembourg Clinical Proteomics Center (LCP), Luxembourg Institute of Health , Strassen 1445, Luxembourg.|University of Luxembourg , Esch-sur-Alzette 4365, Luxembourg.
authors.forename: Bruno
authors.initials: B
authors.lastname: Domon
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D006801:Humans; D008137:Longitudinal Studies; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D059748:Proteolysis; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D012015:Reference Standards; D013026:Space Flight; D053719:Tandem Mass Spectrometry; D016482:Urinalysis / Q000379:methods / Q000592:standards
chemical_list: D010446:Peptide Fragments; D020543:Proteome
keywords: Targeted proteomics; normal protein concentration; quantification; spectral library; urine