Fractionation of liver proteins by preparative electrophoresis

被引:12
作者
Fountoulakis, M
Juranville, JF
Tsangaris, G
Suter, L
机构
[1] F Hoffmann La Roche & Co Ltd, Ctr Med Genom, CH-4070 Basel, Switzerland
[2] Acad Athens, Fdn Biomed Res, Athens, Greece
[3] F Hoffmann La Roche & Co Ltd, Drug Safety, CH-4002 Basel, Switzerland
关键词
proteomics; preparative electrophoresis; liver; low-abundance proteins; mass spectrometry;
D O I
10.1007/s00726-003-0041-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Proteomics offers unique possibilities to investigate changes in the levels and modifications of proteins involved in the pathomechanisms of diseases and toxic events. However, search for potential drug targets and disease or toxicity markers is limited by the fact that mainly the high-abundance, hydrophilic proteins are visualized in two-dimensional gels. Here we studied the enrichment of rat liver cytosolic proteins by preparative electrophoresis. Preparative electrophoresis was performed with the PrepCell apparatus in the presence of 0.1% lithium dodecyl sulfate. Lithium dodecyl sulfate was exchanged against agents compatible with isoelectric focusing prior to the two-dimensional gel electrophoresis. Proteins were identified from two-dimensional gels by matrix-assisted laser desorption ionization time-of-flight mass specrometry. Low- and middle-size proteins and low-abundance proteins, which had not been found before, were enriched by preparative electrophoresis. The present study represents a contribution of proteomics in the quantification of differences in the levels of low-abundance liver proteins in toxicity studies.
引用
收藏
页码:27 / 36
页数:10
相关论文
共 44 条
[1]  
Berndt P, 1999, ELECTROPHORESIS, V20, P3521, DOI 10.1002/(SICI)1522-2683(19991201)20:18<3521::AID-ELPS3521>3.0.CO
[2]  
2-8
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   The protective effect of taurine against thioacetamide hepatotoxicity of rats [J].
Dogru-Abbasoglu, S ;
Kanbagli, Ö ;
Balkan, J ;
Çevikbas, U ;
Aykaç-Toker, G ;
Uysal, M .
HUMAN & EXPERIMENTAL TOXICOLOGY, 2001, 20 (01) :23-27
[5]   Protein expression in Down syndrome brain [J].
Engidawork, E ;
Lubec, G .
AMINO ACIDS, 2001, 21 (04) :331-361
[6]   Challenges and limitations of gene expression profiling in mechanistic and predictive toxicology [J].
Fielden, MR ;
Zacharewski, TR .
TOXICOLOGICAL SCIENCES, 2001, 60 (01) :6-10
[7]  
Fountoulakis M, 2000, ELECTROPHORESIS, V21, P2148, DOI 10.1002/1522-2683(20000601)21:11<2148::AID-ELPS2148>3.0.CO
[8]  
2-X
[9]   Reference map of the low molecular mass proteins of Haemophilus influenzae [J].
Fountoulakis, M ;
Juranville, JF ;
Röder, D ;
Evers, S ;
Berndt, P ;
Langen, H .
ELECTROPHORESIS, 1998, 19 (10) :1819-1827
[10]   PURIFICATION OF INTERFERON GAMMA-INTERFERON GAMMA-RECEPTOR COMPLEXES BY PREPARATIVE ELECTROPHORESIS ON NATIVE GELS [J].
FOUNTOULAKIS, M ;
TAKACSDILORENZO, E ;
JURANVILLE, JF ;
MANNEBERG, M .
ANALYTICAL BIOCHEMISTRY, 1993, 208 (02) :270-276