 | Acta Crystallographica Section D Acta Crystallographica Section D BIOLOGICAL CRYSTALLOGRAPHY |
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![[Figure 3]](rr5103fig3mag.jpg)
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Figure 3
CpSrtD recognizes and cleaves the LPQTGS sorting motif in vitro. (a) Recombinant CpSrtD was incubated in the absence (lane 1) or presence of Aβ1–16 peptides fused to LPETG (lane 2), LPNTGS (lane 3), LPQTGS (lane 4) or LAETG (lane 5) sorting motifs. (b) Catalytic efficiency of recombinant CpSrtD towards Aβ1–16-LPQTGS substrate was measured at room temperature (RT, lane 2), 303 K (lane 3), 310 K (lane 4), 316 K (lane 5) or 323 K (lane 6). (c) CpSrtD was pre-incubated in the absence (lane 1) or presence of EDTA at a concentration of 1 mM (lane 2), 2 mM (lane 3), 5 mM (lane 4), 10 mM (lane 5), 20 mM (lane 6), 50 mM (lane 7), 100 mM (lane 8) or 200 mM (lane 9). EDTA-treated CpSrtD was then incubated with Aβ1–16-LPQTGS and the ability to form the thioacyl intermediate was measured. (d) EDTA-treated (100 mM) CpSrtD was incubated with Aβ1–16-LPQTGS in the absence of metal ions (lane 1) or the presence of 10 mM Ca2+ (lane 2), Cu2+ (lane 3), Co2+ (lane 4), Fe3+ (lane 5), Mg2+ (lane 6), Mn2+ (lane 7), Ni2+ (lane 8) or Zn2+ (lane 9). The formation of thioacyl intermediate in (a), (b) and (c) was analysed by Western blot using mouse α-Aβ (WO2) antibody (top panels), and equal loading was assessed using mouse α-His5 antibody (bottom panels). |
![[Figure 3]](rr5103fig3.jpg)
| BIOLOGICAL CRYSTALLOGRAPHY |
ISSN: 1399-0047
