Displacement affinity chromatography of protein phosphatase one (PP1) complexes
GBG Moorhead, L Trinkle-Mulcahy, M Nimick… - BMC biochemistry, 2008 - Springer
BMC biochemistry, 2008•Springer
Background Protein phosphatase one (PP1) is a ubiquitously expressed, highly conserved
protein phosphatase that dephosphorylates target protein serine and threonine residues.
PP1 is localized to its site of action by interacting with targeting or regulatory proteins, a
majority of which contains a primary docking site referred to as the RVXF/W motif. Results
We demonstrate that a peptide based on the RVXF/W motif can effectively displace PP1
bound proteins from PP1 retained on the phosphatase affinity matrix microcystin-Sepharose …
protein phosphatase that dephosphorylates target protein serine and threonine residues.
PP1 is localized to its site of action by interacting with targeting or regulatory proteins, a
majority of which contains a primary docking site referred to as the RVXF/W motif. Results
We demonstrate that a peptide based on the RVXF/W motif can effectively displace PP1
bound proteins from PP1 retained on the phosphatase affinity matrix microcystin-Sepharose …
Background
Protein phosphatase one (PP1) is a ubiquitously expressed, highly conserved protein phosphatase that dephosphorylates target protein serine and threonine residues. PP1 is localized to its site of action by interacting with targeting or regulatory proteins, a majority of which contains a primary docking site referred to as the RVXF/W motif.
Results
We demonstrate that a peptide based on the RVXF/W motif can effectively displace PP1 bound proteins from PP1 retained on the phosphatase affinity matrix microcystin-Sepharose. Subsequent co-immunoprecipitation experiments confirmed that each identified binding protein was either a direct PP1 interactor or was in a complex that contains PP1. Our results have linked PP1 to numerous new nuclear functions and proteins, including Ki-67, Rif-1, topoisomerase IIα, several nuclear helicases, NUP153 and the TRRAP complex.
Conclusion
This modification of the microcystin-Sepharose technique offers an effective means of purifying novel PP1 regulatory subunits and associated proteins and provides a simple method to uncover a link between PP1 and additional cellular processes.
Springer
