|RPN2 and DDOST slightly accumulate in endogenous VAPB immunoprecipitates upon proteasome inhibition|
|Protein name||UniProt ID||MW (Da)||SILAC ratio L/Ha|
|L + MG 2 hr||H + MG 2 hr||L + MG 6 hr||H + MG 6 hr|
|RPN2||P04844||69,284||1.11 ± 0.02 (4)||0.88 ± 0.00 (2)||1.30 ± 0.10 (4)||0.96 ± 0.02 (6)|
|DDOST||P39656||50,801||-||-||1.12 ± 0.00 (2)||0.64 ± 0.00 (1)|
aLight- (L) or heavy-labeled (H) cells were treated with MG132 (MG) for 2 or 6 hr, as indicated. Equal amounts of light- and heavy-labeled extracts were mixed, and endogenous VAPB was immunoprecipitated using specific antibodies. The light/heavy SILAC ratios (L/H) determined by mass spectrometry are indicated. L + MG indicates that the light-labeled samples were treated with MG132. Proteins whose interaction with VAPB is stimulated by proteasome inhibition accumulate in these samples, resulting in L/H ratios higher than 1. H + MG indicates that the heavy-labeled samples were treated with MG132. Protein accumulation in the heavy-labeled samples results in L/H ratios lower than 1. The L/H for proteins that are not affected by proteasome inhibition will be close to 1. The numbers in parenthesis represent the number of spectra analyzed for each sample.
Baron et al.
Baron et al. BMC Biology 2014 12:39 doi:10.1186/1741-7007-12-39