Quantitative proteome analysis using D-labeled N-ethylmaleimide and 13C-labeled iodoacetanilide by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
A new methodology for quantitative analysis of proteins is described, applying stable-isotope labeling by small organic molecules combined with one- or two-dimensional electrophoresis and MALDI-TOF-MS, also allowing concurrent protein identification by peptide mass fingerprinting. Our method eliminates fundamental problems in other existing isotope-tagging methods requiring liquid chromatography and MS/MS, such as isotope effects, fragmentation, and solubility. It is also anticipated to be more practical and accessible than those LC-dependent methods.
Kurono
Sadamu
S
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA.
Kurono
Tamie
T
Komori
Naoka
N
Niwayama
Satomi
S
Matsumoto
Hiroyuki
H
D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry*; D003903:Deuterium; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005033:Ethylmaleimide / Q000737:chemistry*; D019476:Insect Proteins / Q000032:analysis*; D007460:Iodoacetamide / Q000737:chemistry*; D007553:Isotope Labeling; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D002247:Carbon Isotopes; D019476:Insect Proteins; D020543:Proteome; D003903:Deuterium; D005033:Ethylmaleimide; D007460:Iodoacetamide