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Displacement affinity chromatography of protein phosphatase one (PP1) complexes.
- Source :
-
BMC biochemistry [BMC Biochem] 2008 Nov 10; Vol. 9, pp. 28. Date of Electronic Publication: 2008 Nov 10. - Publication Year :
- 2008
-
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.<br />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.<br />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.
- Subjects :
- Amino Acid Motifs
Animals
Binding Sites
Glycogen isolation & purification
HeLa Cells
Humans
Microcystins chemistry
Microcystins metabolism
Muscle, Skeletal metabolism
Protein Phosphatase 1 chemistry
Protein Subunits chemistry
Protein Subunits metabolism
Rabbits
Sepharose chemistry
Chromatography, Affinity methods
Protein Phosphatase 1 isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1471-2091
- Volume :
- 9
- Database :
- MEDLINE
- Journal :
- BMC biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 19000314
- Full Text :
- https://doi.org/10.1186/1471-2091-9-28