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.
Broedel
Oliver
O
Department of Radiology and Nuclear Medicine, Radiochemistry, Charite Universitatsmedizin, Campus Benjamin Franklin, Berlin, Germany.
Krause
Eberhard
E
Stephanowitz
Heike
H
Schuemann
Michael
M
Eravci
Murat
M
Weist
Stephanie
S
Brunkau
Cindy
C
Wittke
Janosch
J
Eravci
Selda
S
Baumgartner
Andreas
A
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