False positives in MALDI-TOF detection of ERbeta in mitochondria.
Recently, Yang et al. reported that estrogen receptor beta (ERbeta) is a mitochondrial protein rather than a nuclear receptor. Because this claim would lead to a significant change in our understanding of estrogen signaling, we have attempted to reproduce the MALDI-TOF data of Yang et al. We separated proteins extracted from mouse liver mitochondria by SDS-PAGE and analysed a gel band covering the molecular weight range of 50-65 kDa by MALDI-TOF/TOF. Analysis of the data with the MASCOT database algorithm provided no evidence for the presence of ERbeta in the mitochondria. If we search (as the authors did) with only the peptide masses which match to tryptic fragments of ERbeta, ERbeta is identified with a significant score of 69. However, fragmentation of these peptides shows that they are not from ERbeta. Our conclusion is that ERbeta cannot be identified by MALDI-TOF from a mixture of mitochondrial proteins resolved on SDS-PAGE.
Schwend
Thomas
T
Department of Biosciences and Nutrition, Karolinska Institutet/NOVUM, Hälsovägen 7, S-141 86 Huddinge, Sweden.
Gustafsson
Jan-Ake
JA
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D047629:Estrogen Receptor beta / Q000032:analysis*; D051379:Mice; D024101:Mitochondrial Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D047629:Estrogen Receptor beta; D024101:Mitochondrial Proteins