Protein disulphide isomerase family members show distinct substrate specificity: P5 is targeted to BiP client proteins -- Jessop et al. 122 (23): 4287 Data Supplement - JCS059154 Supplementary Material -- Journal of Cell Science

Protein disulphide isomerase family members show distinct substrate specificity: P5 is targeted to BiP client proteins
J Cell Sci Jessop et al. 122: 4287

JCS059154 Supplementary Material

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Supplemental Figure S1 -
Fig. S1. Stable cell lines express wild-type and substrate-trapping mutants of PDI family members at equivalent levels. Total cell lysates from each of the wild-type (WT) or substrate trapping mutants (mut) cell lines were separated on a 7.5-12.5% gradient gel and PDI family members identified by western blotting using a V5 antibody. Cell lines are as indicated.
Supplemental Figure S2 -
Fig. S2. Wild-type PDI forms a mixed disulphide with Ero1. The stable cell line expressing wild-type PDI was lysed and V5-tagged PDI immunoisolated with V5 agarose. Immunoisolated material was eluted in SDS-PAGE sample buffer and separated on a 10% gel under either reduced (red, lane 1) or non-reducing (non-red, lane 2) conditions. The presence of Ero1α in the immunoisolated material was verified by western blotting with antibody to Ero1α. The bands for immunoglobulin heavy chain is indicated (IgG) as are the bands for Ero1α and Ero1α and PDI mixed disulphide.
Supplemental Figure S3 -
Fig. S3. Folding time course of immunoglobulin heavy chain. Translation of immunoglobulin heavy chain was carried out in the presence of SP cells isolated form HT1080 cells for the indicated times. Products of translation were separated under non-reducing conditions through a 7.5% gel and radiolabelled protein visualised by autoradiography.
Supplemental Figure S4 -
Fig. S4. Folding time course of PTX3. The translation of PTX3 was carried out in the presence of SP cells isolated from HT1080 cells for the times indicated. Samples were separated under reducing (red) or non-reducing conditions as indicated and through a 7.5-12.5% gradient gel. Radiolabelled protein was visualised by autoradiography.
Supplemental Figure S5 -
Fig.S5. Folding time course of procollagen C-propeptide. The translation of procollagen C-propeptide was carried out in the presence of SP cells isolated from HT1080 cells for the times indicated. Samples were separated under reducing (red) or non-reducing conditions as indicated and through a 7.5-12.5% gradient gel. Radiolabelled protein was visualised by phosphorimage analysis. The mobility of the monomer, dimer and trimer of the C-propeptide are as indicated.