Maturation kinetics of a multiprotein complex revealed by metabolic labeling

The project aims at computing in vivo co-assembly kinetics from metabolomic-inspired approaches. In short, we performed reciprocal pull-down experiments from complex cellular components at different time points following pulsed-SILAC metabolic labeling. We applied the method to 320 pairs of yeast nucleoporin (NUP) proteins constituting the ~ 50 MDa Nuclear Pore Complex (NPC). Our analysis reveals a hierarchical principle of the NPC biogenesis where individual subcomplexes form on a minute time-scale, followed by their co-assembly from central to peripheral subunits in a ~ 1 hour long maturation process.