Saposin A Nanoparticles for PANX1 Ligand Binding

Human saposin A was recombinantly produced and purified as previously described^{3 (link),6}. Pure saposin A in HNG buffer (50 mM HEPES at pH 7.5, 150 mM NaCl, 5% (v/v) glycerol) was stored at − 80 °C. saposin A empty nanoparticles (Salipro-Empty particles) were prepared as previously described^{3 (link)}. PreScission protease was purchased from Cytiva. PANX1 ligands for SPR binding measurements were Benzoylbenzoyl-ATP (Sigma-Aldrich, CAS 112898-15-4), Carbenoxolone (sourced at AstraZeneca) and Spironolactone (sourced at AstraZeneca).

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Drulyte I., Gutgsell A.R., Lloris-Garcerá P., Liss M., Geschwindner S., Radjainia M., Frauenfeld J, & Löving R. (2023). Direct cell extraction of membrane proteins for structure–function analysis. Scientific Reports, 13, 1420.

Publication 2023

PreScission protease Benzoylbenzoyl-ATP

Benzoylbenzoyl atp Buffer Carbenoxolone Glycerol Hepes Human Ligands Nacl Protease Saposin a Saposin c Spironolactone

Corresponding Organization :

Other organizations : Thermo Fisher Scientific (Netherlands), AstraZeneca (Sweden), Active Biotech (Sweden), Thermo Fisher Scientific (Germany)

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Variable analysis

independent variables

Recombinant production and purification of human saposin A
Preparation of saposin A empty nanoparticles (Salipro-Empty particles)

dependent variables

Binding measurements of PANX1 ligands (Benzoylbenzoyl-ATP, Carbenoxolone, Spironolactone) using SPR

control variables

HNG buffer (50 mM HEPES at pH 7.5, 150 mM NaCl, 5% (v/v) glycerol) for storing purified saposin A
PreScission protease purchased from Cytiva

controls

Positive control: None mentioned
Negative control: None mentioned

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