Supplementary Figure 2: Detection of a covalent conjugate and non-covalent complex of Atg12 and Atg5 by mass spectrometry.
From: Evolution from covalent conjugation to non-covalent interaction in the ubiquitin-like ATG12 system
a, Schematic representation of mass spectrometry of Atg12 and Atg5 in K. phaffii and S. cerevisiae. Lysates from K. phaffii and S. cerevisiae cells expressing FLAG-KpAtg12 and FLAG-ScAtg12, respectively, were subjected to immunoprecipitation using anti-FLAG antibody. The gels were stained with CBB and three portions at the positions corresponding to the Atg12 monomer, Atg5 monomer, and Atg12–Atg5 conjugate were excised for in-gel trypsin digestion. Trypsin digests from each sample were labeled by stable isotope dimethyl labeling method. Mixtures of labeled peptides were subjected to LC-MS/MS and peptides were identified and quantified. b, List of sequences and quantities of peptides identified. The number indicates the area of the chromatogram of the peptides identified.
