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Table 2 TRC complex subunits slightly accumulate in endogenous VAPB immunoprecipitates upon proteasome inhibition

From: VAPB/ALS8 interacts with FFAT-like proteins including the p97 cofactor FAF1 and the ASNA1 ATPase

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
ASNA1 O43681 38,793 1.31 ± 0.08 (31) 0.79 ± 0.07 (27) 1.42 ± 0.10 (32) 0.87 ± 0.06 (35)
BAG6 P46379 119,409 1.47 ± 0.18 (36) 0.77 ± 0.11 (36) 1.43 ± 0.19 (52) 0.94 ± 0.10 (43)
GET4 Q7L5D6 36,504 1.39 ± 0.11 (8) 0.73 ± 0.06 (11) 1.30 ± 0.08 (9) 0.84 ± 0.03 (13)
UBL4A P11441 17,777 1.50 ± 0.10 (7) 0.74 ± 0.04 (9) 1.51 ± 0.15 (7) 0.89 ± 0.04 (10)
  1. aLight- (L) and 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.