In vitro study of interaction of 17β-hydroxysteroid dehydrogenase type 10 and cyclophilin D and its potential implications for Alzheimer’s disease

Reagents

NaCl, NaOH, KCl, MgCl₂, CaCl₂, bovine serum albumin (BSA) and all buffers: sodium acetate (SA10; 10 mM, pH 5.0), MES (10 mM, pH 5.0), HEPES (10 mM), high ionic strength phosphate-buffered saline (PBS_Na; 10 mM phosphate, 2.9 mM KCl, 750 mM NaCl, pH 7.4), were purchased from Sigma-Aldrich, Czech Republic. Oligo-ethylene glycol thiols 11-mercapto-hexa(ethyleneglycol)undecyloxy acetic acid (HS-C₁₁-(EG)₆-OCH₂-COOH) and 11-Mercapto-tetra(ethyleneglycol)undecanol (HS-C₁₁-(EG)₄-OH) were purchased from Prochimia, Poland. Ethanolamine hydrochloride (EA), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were purchased from Biacore, Sweden. All buffers were prepared using deionized Milli-Q water (Merck, Czech Republic). 17β-HSD10 (human, recombinant), cypD (human, recombinant) and an antibody against cypD (Ab(cypD)) were…