Displacement affinity chromatography of protein phosphatase one (PP1) complexes

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Author(s)

  • Greg B.G. Moorhead
  • Laura Trinkle-Mulcahy
  • Mhairi Nimick
  • Veerle De Wever
  • David G. Campbell
  • Robert Gourlay
  • Angus I. Lamond

Detail(s)

Original languageEnglish
Article number28
Journal / PublicationBMC Biochemistry
Volume9
Issue number1
Publication statusPublished - 2008
Externally publishedYes

Link(s)

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 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. © 2008 Moorhead et al; licensee BioMed Central Ltd.

Research Area(s)

Citation Format(s)

Displacement affinity chromatography of protein phosphatase one (PP1) complexes. / Moorhead, Greg B.G.; Trinkle-Mulcahy, Laura; Nimick, Mhairi; De Wever, Veerle; Campbell, David G.; Gourlay, Robert; Lam, Yun Wah; Lamond, Angus I.

In: BMC Biochemistry, Vol. 9, No. 1, 28, 2008.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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