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Tytuł:
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Proteomic analysis of normal human urinary proteins isolated by acetone precipitation or ultracentrifugation.
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Autorzy:
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Thongboonkerd V; Core Proteomics Laboratory and Molecular Signaling Group, Kidney Disease Program, Department of Medicine, University of Louisville, 570 South Preston Street, Louisville, KY 40202, USA.
McLeish KR
Arthur JM
Klein JB
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Źródło:
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Kidney international [Kidney Int] 2002 Oct; Vol. 62 (4), pp. 1461-9.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.
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Język:
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English
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Imprint Name(s):
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Publication: 2016- : New York : Elsevier
Original Publication: New York, Springer-Verlag.
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MeSH Terms:
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Proteomics*
Proteins/*analysis
Urine/*chemistry
Acetone ; Chemical Precipitation ; Computational Biology ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Peptide Mapping ; Serine Endopeptidases ; Solvents ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Staining and Labeling ; Tryptases ; Ultracentrifugation
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Grant Information:
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R01 HL 66358-01 United States HL NHLBI NIH HHS
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Substance Nomenclature:
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0 (Proteins)
0 (Solvents)
1364PS73AF (Acetone)
EC 3.4.21.- (Serine Endopeptidases)
EC 3.4.21.59 (Tryptases)
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Entry Date(s):
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Date Created: 20020918 Date Completed: 20030225 Latest Revision: 20220408
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Update Code:
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20240104
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DOI:
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10.1111/j.1523-1755.2002.kid565.x
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PMID:
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12234320
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Background: Proteomic techniques have recently become available for large-scale protein analysis. The utility of these techniques in identification of urinary proteins is poorly defined. We constructed a proteome map of normal human urine as a reference protein database by using two differential fractionated techniques to isolate the proteins.
Methods: Proteins were isolated from urine obtained from normal human volunteers by acetone precipitation or ultracentrifugation, separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and identified by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry followed by peptide mass fingerprinting.
Results: A total of 67 protein forms of 47 unique proteins were identified, including transporters, adhesion molecules, complement, chaperones, receptors, enzymes, serpins, cell signaling proteins and matrix proteins. Acetone precipitated more acidic and hydrophilic proteins, whereas ultracentrifugation fractionated more basic, hydrophobic, and membrane proteins. Bioinformatic analysis predicted glycosylation to be the most common explanation for multiple forms of the same protein.
Conclusions: Combining two differential isolation techniques magnified protein identification from human urine. Proteomic analysis of urinary proteins is a promising tool to study renal physiology and pathophysiology and to determine biomarkers of renal disease.