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BMC Biochem. 2008 Nov 10;9:28. doi: 10.1186/1471-2091-9-28.

Displacement affinity chromatography of protein phosphatase one (PP1) complexes.

Author information

1
Department of Biological Sciences, University of Calgary, 2500 University Dr, NW Calgary, AB T2N 1N4, Canada. moorhead@ucalgary.ca

Abstract

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 IIalpha, 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.

PMID:
19000314
PMCID:
PMC2587467
DOI:
10.1186/1471-2091-9-28
[Indexed for MEDLINE]
Free PMC Article
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