Figure 2.
Illustration of sample preparation work flow for large-scale relative quantification of differentially cross-linked E. coli cells. Two batches of cells were cultured for biological replicates. Cultured cells from one batch were split into two equal halves and cross-linked with d0-BDP-NHP and d8-BDP-NHP respectively. After cell lysis, 8 mg of cross-linked E. coli lysates were pooled in d0/d8 1:2 and 2:1 ratios based on total protein mass. The lysates were digested, fractioned by SCX, enriched with avidin beads, and then analyzed by LC-ReACT in triplicates for MS1 peak intensity based quantification.