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Displacement affinity chromatography of protein phosphatase one (PP1) complexes
- Source :
- BMC Biochemistry, Vol 9, Iss 1, p 28 (2008), BMC Biochemistry
- Publication Year :
- 2008
- Publisher :
- BMC, 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. 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.
- Subjects :
- Binding Sites
animal structures
Microcystins
Sepharose
Methodology Article
Amino Acid Motifs
fungi
lcsh:Animal biochemistry
macromolecular substances
Biochemistry
environment and public health
Chromatography, Affinity
lcsh:Biochemistry
enzymes and coenzymes (carbohydrates)
Protein Subunits
Protein Phosphatase 1
Animals
Humans
lcsh:QD415-436
Rabbits
Muscle, Skeletal
Molecular Biology
lcsh:QP501-801
Glycogen
HeLa Cells
Subjects
Details
- Language :
- English
- ISSN :
- 14712091
- Volume :
- 9
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- BMC Biochemistry
- Accession number :
- edsair.pmid.dedup....5f3e8332d4f0e5c9f30f395d843446cf