Proteins of rat serum: I. Establishing a reference two-dimensional electrophoresis map by immunodetection and microbore high performance liquid chromatography-electrospray mass spectrometry.
In the present investigation, we have identified 56 major spots, or spot rows, corresponding to 22 proteins, in the 2-DE pattern of adult male rats. This was done mainly by applying two complementary techniques, namely immunoblotting and high performance liquid chromatography-mass spectrometry (HPLC-MS) peptide mapping. Glycoproteins were characterized by affinity blotting with six lectins. We have also detailed how rat serum differs from human serum in two main respects: (i) relative abundance of individual proteins, which amounts in some cases to a complete absence in either sample, and (ii) varying molecular parameters for homologous proteins. It was thus possible to establish a first-generation reference map of rat serum proteins, which can be accessed through http://weber.u.washington.edu/ruedilab/aebersold++ +.html. We hope the present database will be a useful reference for the evaluation of changes in serum protein distribution in the course of pharmacological and toxicological studies. The recognition of species-specific proteins appears of special relevance in this respect.
Department of Molecular Biotechnology, University of Washington, School of Medicine, Seattle, USA.
P
P
Haynes
I
I
Miller
R
R
Aebersold
M
M
Gemeiner
I
I
Eberini
M R
MR
Lovati
C
C
Manzoni
M
M
Vignati
E
E
Gianazza
D000818:Animals; D001798:Blood Proteins / Q000032:analysis / Q000592:standards*; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D015151:Immunoblotting; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D010449:Peptide Mapping / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards
D001798:Blood Proteins