MPM-2 Mediated Immunoprecipitation of Proteins Undergoing Proline-directed Phosphorylation

MPM-2 Mediated Immunoprecipitation of Proteins Undergoing Proline-directed Phosphorylation

Blog Article

Immunoprecipitation (IP) represents a widely utilized biochemical method to isolate a specific protein from a complex mixture taking advantage of an antibody that specifically recognizes that particular target molecule.This procedure is extremely versatile and can be applied to concentrate a specific protein, to identify interacting partners in complex with it or to detect post-translational modifications.The Right Seat Adjustment Lever mitotic protein monoclonal 2 (MPM-2) is an antibody originally raised against extracts of synchronized mitotic HeLa cells to identify proteins selectively present in mitotic, and not in interphase-cells (Davis et al.

, 1983).MPM-2 recognizes phosphorylated serine or threonine residues followed by proline (pS/T-P), consensus epitopes generated by the concerted action of proline-directed protein kinases and phosphatases (Lu et al., 2002).

These reversible phosphorylation events have emerged to control various cellular processes by promoting conformational changes on Course a pied - Homme - Vetements - Manteau - Impermeable the target that are not simply due to the phosphorylation event per se.These motifs, once phosphorylated, are able to recruit Pin1 (Peptidyl-prolyl Isomerase NIMA interacting protein 1) (Lu et al., 1996; Lu and Zhou, 2007), a chaperone which drives the cis/trans isomerization reaction on the peptide bond, switching the substrate between functionally diverse conformations (Lu, 2004; Wulf et al.

, 2005).This protocol describes a general MPM-2 based immunoprecipitation strategy using the scaffolding molecule postsynaptic density protein-95 (PSD-95) (Chen et al., 2005), a neuronal Pin1 target (Antonelli et al.

, 2016), as an example to illustrate the detailed procedure.