Table 2.
Summary of the hetero-oligomerization properties of HSPB6 deletions and point mutations with HSPB1
| 10 amino acid deletions |
5 amino acid deletions |
Point mutations |
||||||
|---|---|---|---|---|---|---|---|---|
| Construct | SECa | S–Sb | Construct | SECa | S–Sb | Constructc | SECa | S–Sb |
| ΔN11 | WT | 1 | Δ21–25 | WT | 1 | 10swap | WT | 1 |
| Δ11–20 | WT | 1 | Δ26–30 | Remnant HSPB6 | 1:0.5:1 | 5swap | Remnant HSPB6 | 1:0.5:1 |
| Δ21–30 | Remnant HSPB6 | 1:0.5:1 | Δ31–35 | Remnant HSPB6 | 1:0.5:1 | R32A | WT | 1 |
| Δ31–40 | Remnant HSPB6 | 1:0.5:1 | Δ36–40 | Remnant HSPB6 | 1:2:1 | F33A | Remnant HSPB6 | 1:0.5:1 |
| Δ41–50 | WT | 1 | Δ51–55 | Small | 1 | FFAA | Remnant HSPB6 | 1:0.5:1 |
| Δ51–60 | Small | 1 | Δ56–60 | WT | 1 | |||
| Δ61–70 | WT | 1 | ||||||
a SEC profile of each HSPB6 construct when mixed in equimolar amounts with HSPB1 and heated are shown. WT, closely resembling the mixture of WT HSPB6 and HSPB1. Remnant HSPB6, chromatograms contain an additional late eluting peak corresponding to the HSPB6 dimer position. Small, chromatograms show a profile biased to an approximately 140-kDa species.
b Ratio of disulfide cross-linked HSPB1 homodimer, HSPB1 and HSPB6 construct heterodimer, and HSPB6 construct homodimer as evaluated by non-reducing SDS-PAGE. A single value corresponds to heterodimers only.
c 10swap corresponds to the HSPB6 G26S/R32A/E35L mutant. The 5swap corresponds to the HSPB6 G36P/RL37R/E39P/A40E mutant. FFAA corresponds to the HSPB6 F29A/F33A mutant.