Database searching artifacts

Fig. 18 The new strategy outlined in Fig. 17 was evaluated in the analysis of protein-affinity pull-down experiments, to distinguish specifically enriched sample components from experimental artifacts. The carboxy terminus of Hsp70-interacting protein (CHIP) was expressed as GST fusion protein in E. coli and used as bait to affinity purify potential interaction partners from a crude human brain protein extract, a Peptide profile obtained from the LC-MALDl MS analysis of the protein isolate obtained from the affinity pull-down experiment. Blue dots-, unpaired peptides, red dots paired signals, black dots all detected monoisotopic peaks, b Mass spectrum acquired from fraction 89. The signal of m/z 1253.62 is, apart from the internal calibrants, the only unpaired signal in the spectrum, indicating that it originates from a protein that specifically interacts with CHIP, c MS/MS spectrum of m/z 1253.62. A database search identified the peptide FEEL-NADLFR of heat shock cognate 71-kDa protein. Reproduced from [316] with permission from American Chemical Society, 2006...

$Fig. 18 The new strategy outlined in Fig. 17 was evaluated in the analysis of protein-affinity pull-down experiments, to distinguish specifically enriched sample components from experimental artifacts. The carboxy terminus of Hsp70-interacting protein (CHIP) was expressed as GST fusion protein in E. coli and used as bait to affinity purify potential interaction partners from a crude human brain protein extract, a Peptide profile obtained from the LC-MALDl MS analysis of the protein isolate obtained from the affinity pull-down experiment. Blue dots-, unpaired peptides, red dots paired signals, black dots all detected monoisotopic peaks, b Mass spectrum acquired from fraction 89. The signal of m/z 1253.62 is, apart from the internal calibrants, the only unpaired signal in the spectrum, indicating that it originates from a protein that specifically interacts with CHIP, c MS/MS spectrum of m/z 1253.62. A database search identified the peptide FEEL-NADLFR of heat shock cognate 71-kDa protein. Reproduced from [316] with permission from American Chemical Society, 2006...$

Initially, by defining key-words aiming at limiting the search to the rather specific theme under study, focusing on words such as, Eye blink , EEG , Blink , Artifact , Rate , Fatigue , Frequency , Blink frequency , Video , Image capture , Environmental conditions , Mental load , Temperature , Relative Humidity , which in groups in the databases with the appropriate Boolean operator AND . This way a total... [Pg.231]

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