Table 2.
Expression of soluble domains of newly introduced targets
| Target gene/Uniprot ID | Soluble constructs/tested |
Domains produced (aa range) | Prior status | |||
|---|---|---|---|---|---|---|
| N-His | C-His | His/Trx | Baculo | |||
| AMPH/P49418 |
5/7 |
5/8 |
4/8 |
8/8 |
BAR domain (aa34–236) |
67% identity to human Bin1 [21] |
| CNO/Q9NUP1 |
2/7 |
0/9 |
5/7 |
3/7 |
(aa1–217) |
None reported |
| ETV1/P50549 |
5/8 |
0 |
7/7 |
7/7 |
PEA3, ETS-N-terminal domain (aa223–326) |
Short homology to ets-1 |
| HCK (SH2-SH3)/P08631 |
6/8 |
2/8 |
5/8 |
4/7 |
SH3 domain (aa72–138) |
Structure solved (PDB:3NHN) [22] |
| HMHA1/Q8IYN3 |
3/13 |
(weak) |
1/12 |
5/13 |
FCH: Fes/CIP4 homology domain (aa254–521) |
31% homology in RhoGAP domain |
| INADL/Q8NI35 |
3/5 |
1/6 |
2/5 |
4/5 |
PDZ 2 + 3 (aa254–530) |
PDZ domain structures solved by NMR (PDB 2DB5, 2DMZ and 2DAZ) |
| MSX2/P35548 |
5/8 |
2/9 |
7/8 |
3/8 |
Homeobox domain (aa143–211) |
Highly homologous to MSX1 in homeobox |
| RBM3/P98179 |
6/6 |
5/6 |
6/6 |
6/6 |
RRM1 (aa1–106) |
68% identity to closest PDB homologue 1X5S |
| RCC1/P18754 |
0/1 |
1/1 |
1/1 |
1/1 |
Full-length RCC1 (aa1–421) |
Structure solved (PDB:1A12) [23] |
| TEX9/Q8N6V9 |
0/3 |
2/4 |
(weak) |
(weak) |
(aa172–391) |
None reported |
| PLEKHH1/Q9ULM0 |
0/16 |
0/16 |
4/16 |
ND |
Refolded MyTH4 domain (aa832–990) |
None reported |
| HCK (kinase)/P08631 |
0/4 |
ND |
5/6 |
(weak) |
SH3_SH2_Kinase (aa72–526) |
Structure solved (PDB:1QCF); expressed in mammalian cells [24] |
| Secreted proteins | ||||
|---|---|---|---|---|
| Target gene | E. coli | Baculo | ||
| SPON1/Q9HCB6 |
1/3 |
1/6 |
Spondin_N domain(aa194–413) |
Structure solved (aa1–200), PDB:3COO [25] |
| CST3/P01034 |
1/1 |
1/1 |
Cystatin domain (aa27–146) |
Structure solved (PDB: 1G96). Periplasmic expression in E. coli[26] |
| FAT3/Q8TDW7-3 |
11/24 |
ND |
Cadherin 32 (aa3443–3553); EGF_like (aa4016–4137) |
None |
| ITGB5/P18084 | ND | (weak) | ND | 55% identical to integrin beta-3 structure (PDB:3IJE), expressed as complex with ITGAV in insect or mammalian cells [27,28] |
For each target and vector, the number of constructs expressing soluble proteins is listed, out of the number of constructs tested. ‘Domains produced’ indicates the segment of the target protein included in a construct that was selected for scale-up and purification. N-His, C-His, Trx and Bacolu refer to clones in the vectors pNIC28-Bsa4, pNIC-CTHF, pNH-TrxT and the baculovirus transfer vector pFB-LIC-Bse [4].