Table 3.
Creld2 is part of mixed disulphide complexes with structural proteins, chaperones and PDIs
| Protein | Type | HT1080 | Creld2 WT | Creld2 N-CXXA | Creld2 C-CXXA | Creld2 N/C-CXXA |
|---|---|---|---|---|---|---|
| BiP | Chaperone | 0/0 | 4/0/6 | 16/17/19 | 3/0/9 | 17/16/17 |
| GRP94 | Chaperone | 0/0 | 0/0/3 | 0/11/3 | 0/0/2 | 8/8/9 |
| HSP90β | Chaperone | 4/0 | 4/0/4 | 10/5/10 | 0/0/4 | 17/5/17 |
| HSPA8 | Chaperone | 0/0 | 6/0/3 | 7/13/9 | 2/0/4 | 14/5/14 |
| SERPINH1 | Chaperone | 0/0 | 0/0/0 | 2/6/2 | 0/0/0 | 4/3/3 |
| PDIA1 | PDI | 0/0 | 3/0/0 | 3/6/4 | 0/0/4 | 7/6/5 |
| PDIA3 | PDI | 0/0 | 0/0/0 | 3/6/5 | 0/0/7 | 12/11/8 |
| PDIA4 | PDI | 0/0 | 0/0/0 | 0/3/0 | 0/0/0 | 7/4/4 |
| LAMB3 | ECM protein | 0/0 | 0/0/0 | 11/15/22 | 2/0/14 | 14/13/17 |
| α1(VI) chain | ECM protein | 0/0 | 0/0/0 | 7/9/15 | 0/0/0 | 9/5/5 |
| α3(VI) chain | ECM protein | 0/0 | 0/0/0 | 3/9/22 | 0/0/14 | 19/15/12 |
| TSP1 | ECM protein | 0/0 | 0/0/0 | 14/11/23 | 0/0/5 | 6/0/7 |
| Integrin alpha-3 | Receptor | 0/0 | 0/0/0 | 6/4/14 | 0/0/0 | 9/6/10 |
| Pyruvate kinase isozymes M1/M2 | Glycolytic enzyme | 0/0 | 0/0/9 | 10/11/13 | 0/0/5 | 11/5/16 |
A summary of the proteins identified by spectral counting from V5-precipitated complexes of the Creld2 substrate-trapping mutants. Total protein pools were analysed by LC-MS/MS and the data analysed using Mascot against the UniProt human database and validated with Scaffold using peptide/protein confidence values of 0.95 and 0.99, respectively. Positively identified proteins were defined as those having a number of matched peptide spectra greater than two. Three biological replicates were used in all experiments and the number of spectra from each experiment is separated with a forward slash (/).
Key: N/C-CXXA = amino and carboxyl terminal double substrate-trapping mutant; C-CXXA = carboxyl terminal substrate-trapping mutant; N-CXXA = amino terminal substrate-trapping mutant; WT = wild-type Creld2; HT1080 = untransfected HT1080 cells; PDI = protein disulphide isomerase; PPI = peptidylprolyl isomerase; ECM = extracellular matrix.