Displacement affinity chromatography of protein phosphatase one (PP1) complexes

Additional file 1:

Supplementary Figure 1. Identification of novel rat liver nuclear PP1 binding and complex proteins by displacement affinity chromatography. Protein was extracted from isolated rat liver nuclei, incubated with microcystin-Sepharose, the matrix washed extensively and eluted with GKKRVRWADLE peptide, followed by elution with 3 M NaSCN [31]. GKKRVRWADLE and NaSCN eluted samples were concentrated separately to an equal volume and run on 10% SDS-PAGE and stained with Collodial blue. In a parallel experiment, the bands shown above were excised, trypsin digested and analyzed by mass spectrometry for identification. The top matched identified proteins for each band(s) are indicated to the left of the figure. Additional matches for each excised band and details of protein identifications are in Additional file 2 online.

Format: JPEG Size: 477KB Download file

Additional file 2:

Additional file 2-Supplementary Table 1. Identification of proteins eluted from microcystin-Sepharose using an 'RVRW' peptide (Rattus norvegicus). Proteins were run on SDS-PAGE and stained with Collodial blue. Individual bands were excised, digested with trypsin and proteins identified by Mass Spectrometry. These proteins are shown by gel band excised with and protein name/description, gene accession numbers, gene name and whether they were previously identified has a PP1 targeting subunit (Known PP1 TS).

Format: XLS Size: 55KB Download file

This file can be viewed with: Microsoft Excel Viewer

Additional file 3:

Supplementary Figure 2. Identification of HeLa nuclear PP1 binding and complex proteins by displacement affinity chromatography. The GKKRVRWADLE elution from Figure 2a has been cropped and the top match identified proteins for each band(s) are indicated on the figure. Additional matches for each excised band and details of protein identifications are in Additional file 4 (Supplementary Table 2) online.

Format: JPEG Size: 471KB Download file

Additional file 4:

Supplementary Table 2. Identification of proteins eluted from microcystin-Sepharose using an 'RVRW' peptide (Homo Sapiens). Proteins were run on SDS-PAGE and stained with Collodial blue. Individual bands were excised, digested with trypsin and proteins identified by Mass Spectrometry. These proteins are shown by gel band excised with gene accession numbers, gene name, number of peptide identified by mass spectrometry and whether they were previously identified has a PP1 targeting subunit (Known PP1 TS).

Format: XLS Size: 30KB Download file

This file can be viewed with: Microsoft Excel Viewer