Exploring the hidden human urinary proteome via ligand library beads.
The human urinary proteome has been reassessed and re-evaluated via a novel concentration/equalization technique, exploiting beads coated with hexameric peptide ligand libraries. These beads act by capturing the whole protein spectra contained in the sample, by drastically reducing the level of the most abundant species, while strongly concentrating the more dilute and rare ones. In a control urine sample, 134 unique proteins could be identified. The first bead eluate (in thiourea, urea, and CHAPS) permitted the identification of 317 gene products, whereas the second eluate (in 9 M urea, pH 3.8) allowed the identification of another 95 unique proteins. By eliminating redundancies, a total of 383 unique gene products could be identified in human urines. This represents a major increment as compared to data reported in recent literature. By comparing our data with those reported to the present, an additional 251 proteins could be added to the list, thus bringing the total unique gene products so far identified in human urines to ca. 800 species.
Castagna
Annalisa
A
Department of Clinical and Experimental Medicine, Unit of Internal Medicine, University of Verona, Italy.
Cecconi
Daniela
D
Sennels
Lau
L
Rappsilber
Juri
J
Guerrier
Luc
L
Fortis
Frederic
F
Boschetti
Egisto
E
Lomas
Lee
L
Righetti
Pier Giorgio
PG
D000328:Adult; D002793:Cholic Acids / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005583:Fourier Analysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008024:Ligands; D008297:Male; D013058:Mass Spectrometry; D019151:Peptide Library; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D016622:Silver Staining; D013890:Thiourea / Q000737:chemistry; D014508:Urea / Q000737:chemistry; D014556:Urine / Q000737:chemistry*
D002793:Cholic Acids; D007477:Ions; D008024:Ligands; D019151:Peptide Library; D011506:Proteins; D020543:Proteome; D014508:Urea; D013890:Thiourea; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate