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fill time: 400?ms). For CID experiments, a normalized collision energy of 35%, an activation q of 0.25 and an activation time of 30?ms were used. The target ion value was set to 2,500 and a dynamic exclusion time of 45?s was placed on previously selected precursor ions. MaxQuant (version 1.2.0.18) was used for data processing (Cox and Mann, 2008; Cox et?al., 2011). Peak lists were searched against the IPI human database (version 3.68) containing 87,061 protein entries. Peptides and proteins were identified with a false discovery rate of ALOX15 13C615N2-lysine and 13C615N4-arginine as heavy labeled amino acids. If peptides were assigned to different proteins or multiple isoforms, the respective proteins were reported as a single protein group. For SILAC (stable isotope labeling of amino acids in cell culture)-based quantification of proteins or protein groups, ��razor�� peptides and a minimum ratio count of two were considered. In addition, BMN 673 price ��requantify�� and ��filter labeled amino acids�� were enabled and low-scoring peptides were excluded. For the SILAC-based analysis of MITRAC12 complexes by denaturing gel electrophoresis, the normalized light-over-heavy selleck compound (L/H; replicates 1 and 3) or heavy-over-light (H/L; replicates 2 and 4) ratios were calculated. Ratios were log-transformed, and the mean log10 ratio across all experiments was calculated for each protein. A one-sided t test was performed to determine p values for all proteins quantified in at least two experiments. Proteins considered to specifically enrich with the bait protein MITRAC12 were required to be identified with at least two peptides as well as quantified with a mean ratio of ��15 and a p value of?