BDB - Target Details

Target

Target Name: PCSK9ΔP'

Target ID:

1953

Target Type:

Protein>Others

Target Sequence:

Synonyms:

Target Source:

Homo sapiens (Human):9606

Target Structure:

Comments:

The deleted α-helix version of PCSK9, called PCSK9ΔP'includes an engineered TEV (tobacco etch virus) protease cleavage site (deletion of residues Y166 and R167; insertion of ENLYFQS sequence between R165 and A168). The above procedure was performed and then purified protein was treated with TEV protease (100 µl at 2.5 mg/ml per 10 mg of protein) for 24 h at 4 °C using dialysis (Slidelyzer 10K, Thermo Scientific) versus 0.15 M NaCl, 50 mM Tris, pH 8.0, 10% glycerol and 3 mM reduced glutathione/ 0.3 mM oxidized glutathione. The repurified PCSK9ΔP'protein (Superdex 75 column) was homogenous and had the expected mass, consistent with the loss of the N-terminal sequence 153SIPWNLERITPPR165ENLYFQ (ENLYFQ is TEV cleavage sequence). The new N terminus of the PCSK9 catalytic domain was SA168 (the serine is from the TEV cleavage sequence), which was confirmed by N-terminal sequencing. Therefore, the PCSK9ΔP'construct is missing the first 15 residues of its native N-terminal sequence (153SIPWNLERITPPRYR167).

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