ReCLIP (Reversible Cross-Link Immuno-Precipitation): An Efficient Method for Interrogation of Labile Protein Complexes (original) (raw)
Figure 4
Cross-linkers can be combined to enhance complex recovery.
(a) Average number of distinct peptides identified in 2 LC-MS/MS runs for E-cadherin, α-catenin, β-catenin, and plakoglobin from A431 cells treated with DSP, DTME, or both compounds simultaneously (DSP + DTME). Error bars represent standard error of the mean. Background levels were similar across all conditions. (b) Western blot analysis of E-cadherin levels in lysates (Lysate), p120 eluates (p120), and control IgG eluates (Control) from A431 cells treated with the indicated cross-linkers. (c) Silver stain analysis of total protein recovery from p120 and control IgG eluates from each condition (DSP, DTME, or DSP + DTME). (d–f) Average distinct peptide recovery of 15 additional putative p120 binding partners under each cross-linking condition. Proteins were grouped based on whether more peptides were detected using the combination of DSP and DTME (d), DSP alone (e) or DTME alone (f).