Establishment of a near-standard two-dimensional human urine proteomic map

Establishment of a near-standard two-dimensional human urine proteomic map
4 Nov 2004
Received: 27 April 2004
Jisun Oh 1, Jae-Hoon Pyo 1, Eun-Hyun Jo 1, Sun-Il Hwang 4, Sun-Chul Kang 3, Jae-Hwan Jung 2 5, Eui-Kyun Park 5, Shin-Yoon Kim 5, Je-Yong Choi 2 5, Jinkyu Lim 1 5 *
PROTEOMICS
Volume 4, Issue 11 , Pages 3485 - 3497
Wiley InterScience
1Department Animal Science and Biotechnology
2Department of Biochemistry School of Medicine, Kyungpook National University,Daegu, Korea
3Division of Food, Biological and Chemical Engineering,Daegu University, Kyung-San, Korea
4Department of Cell Biology University of Connecticut, School of Medicine, Farmington, CT, USA
5Skeletal Diseases Genome Research Center Daegu, Korea

email: Jinkyu Lim (jkylim@knu.ac.kr)
*Correspondence to Jinkyu Lim, Department of Animal Science and Biotechnology, Kyungpook University, 1370 Sankyuk Dong, Daegu, 702-701, Korea Fax: +82-53-950-6750
Keywords
Dialysis ? Lyophilization ? Pooled urine ? Urinary protein
Abstract
A proteomic map for human urine on two-dimensional (2-D) gels has been developed. Initial studies demonstrated that the urine proteins prepared by conventional methods showed interference and poor reproducibility in 2-D electrophoresis (2-DE). To address this issue, urine samples were dialyzed to remove any interfering molecules. The dialysis of urine proteins and the concentration by lyophilization without fractionation significantly improved the reproducibility and resolution and likely represents the total urine proteins on a 2-D gel. In addition, removing albumin from urine using Affi-Gel Blue helped to identify the low-abundant proteins. Using the developed method, we prepared proteins from urine collected from healthy females and males. The large inter- and intra-subject variation in protein profiles on 2-D gels made it difficult to establish a normal human urine proteomic 2-D map. To resolve this problem, urinary proteins were prepared from the pooled urine collected from 20 healthy females and males, respectively. The established male and female urine proteomes separated on 2-D gels were almost identical except for some potential sex-dependent protein spots. We have annotated 113 different proteins on the 2-D gel by peptide mass fingerprinting (PMF). We propose that the established total urine proteome can be used for 2-DE analysis
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