Journal: bioRxiv

Article Title: Substrate-induced modulation of protein-protein interactions within human mitochondrial cytochrome P450-dependent system

doi: 10.1101/2020.11.13.381095

Figure Lengend Snippet: Sensorgrams of interaction CYP – Adx at 25°C in the HBS-EP+ buffer. CYP’s concentration: a – 25 nM, b – 50 nM, c – 125 nM, d – 250 nM. 1 – CYP11A1 – Adx; 2 – CYP11A1 – Adx in the presence of 100 μM cholesterol; 3 – CYP11A1 – Adx in the presence of 100 μM pregnenolone; 4 – CYP11B1 – Adx; 5 – CYP11B1 – Adx in the presence of 100 μM 11-deoxycorticosterone; 6 – CYP11B1 – Adx in the presence of 100 μM 11-deoxycortisol; 7 – CYP11B2 – Adx; 8 – CYP11B2 – Adx in the presence of 100 μM 11-deoxycorticosterone; 9 – CYP11B2 – Adx in the presence of 100 μM 11-deoxycortisol. Arrows indicate the start and the end point of CYP sample’s injections. Double column

Article Snippet: Special reagents for biosensor measurements were obtained from GE Healthcare (GE Healthcare, USA): HBS-EP+ buffer (150 mM NaCl, 10 mM HEPES, 3 mM EDTA, 0.05% P-20 surfactant, pH 7.4), HBS-N buffer (150 mM NaCl, 10 mM HEPES, pH 7.4), 50 mM NaOH solution, 70% glycerol solution, 10 mM acetate buffer (pH 4.5), the amine-coupling kit for covalent immobilization of proteins on the CM4 optical chip: 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-HCl (EDC), N-hydroxysuccinimide (NHS), 1M ethanolamine-HCl (pH 8.5).

Techniques: Concentration Assay