In-Gel 18O labeling for improved identification of proteins from 2-DE Gel spots in comparative proteomic experiments.
The reliability of 2-DE gel-based comparative proteomics is severely impaired by the potential presence of overlapping proteins. We describe a methodological procedure which may solve this problem. Corresponding protein spots from two experimental groups are digested in the presence of 16O and 18O, respectively. Samples are pooled and proteins identified by MS. The 18O/16O-ratios of the different proteins found in the same spot distinguish proteins with altered from those whose intensity is unchanged.
Department of Radiology and Nuclear Medicine, Radiochemistry, Charite Universitatsmedizin, Campus Benjamin Franklin, Berlin, Germany.
Oliver
O
Broedel
Eberhard
E
Krause
Heike
H
Stephanowitz
Michael
M
Schuemann
Murat
M
Eravci
Stephanie
S
Weist
Cindy
C
Brunkau
Janosch
J
Wittke
Selda
S
Eravci
Andreas
A
Baumgartner
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D010103:Oxygen Isotopes / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D010103:Oxygen Isotopes; D011506:Proteins; D020543:Proteome