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  • Interaction of pyridilethanol(phenylethyl)amine derivatives with bovine CYP51
    Zelenko, Urška, biokemik ...
    Sterol 14a-demethylase (CYP51) is a crucial enzyme in the sterol biosynthesis pathway, which catalyzes the oxidative removal of the 14a-methyl group from sterol precursors. Ste rol biosynthesis is an ... essential metabolic pathway producing ergosterol in fungi, phytosterol in plants, and cholesterol in animals. Inhibition of CYP51 activity is lethai in unicellular organisms, influences growth and development in plants, and lowers endogenous cholesterol production in animals. The essential nature of sterol biosynthesis makes CYP51 an important drug target. Currently we are investigating binding of cholesterol biosynthesis inhibitors, a group of pyridilethanol(phenylethyl)amine derivatives, to bovine CYP51 (bCYP51). First, we expressed soluble bCYP51 in E. cali. By changing the strain we substantially increased expression level. bCYP51 was purified first by Ni-NTA affinity chromatography and then by cation exchange chromatography. We have evaluated binding affinities of the selected inhibitors by spectroscopic titration to the purified bCYP51. The interactions between the inhibitors and bCYP51 have been examined using the Iigand-based NMR methods. We have investigated conformations of the bound inhibitors by transferred NOESY experiments. The STD NMR experiments have been used to identify binding of the inhibitors to bCYP51 and to determine the inhibitor enzyme contacts. In viva bCYP51 is membrane-bound microsomal enzyme therefore we have also studied interactions of the inhibitors with micelles by solution NMR spectroscopy.
    Vir: Biochemistry, Biophysics and Structure : conference and abstract book (Str. 77)
    Vrsta gradiva - prispevek na konferenci
    Leto - 2011
    Jezik - angleški
    COBISS.SI-ID - 4717850

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