3L4D

Crystal structure of sterol 14-alpha demethylase (CYP51) from Leishmania infantum in complex with fluconazole

Summary for 3L4D

• Entry DOI: 10.2210/pdb3l4d/pdb
• Descriptor: Sterol 14-alpha demethylase, PROTOPORPHYRIN IX CONTAINING FE, 2-(2,4-DIFLUOROPHENYL)-1,3-DI(1H-1,2,4-TRIAZOL-1-YL)PROPAN-2-OL, ... (5 entities in total)
• Functional Keywords: sterol 14-alpha demethylase, cyp51, p450, heme, oxidoreductase, monooxygenase, endoplasmic reticulum, transmembrane protein, sterol biosynthesis, obtusifoliol, lipids, membrane, iron
• Biological source: Leishmania infantum
• Total number of polymer chains: 4
• Total formula weight: 208795.23

Authors

Lepesheva, G.I.,Hargrove, T.Y.,Wawrzak, Z.,Waterman, M.R. (deposition date: 2009-12-19, release date: 2009-12-29, Last modification date: 2023-09-06)

Primary citation

Hargrove, T.Y.,Wawrzak, Z.,Liu, J.,Nes, W.D.,Waterman, M.R.,Lepesheva, G.I.
Substrate Preferences and Catalytic Parameters Determined by Structural Characteristics of Sterol 14{alpha}-Demethylase (CYP51) from Leishmania infantum.
J.Biol.Chem., 286:26838-26848, 2011

PubMed Abstract: Leishmaniasis is a major health problem that affects populations of ∼90 countries worldwide, with no vaccine and only a few moderately effective drugs. Here we report the structure/function characterization of sterol 14α-demethylase (CYP51) from Leishmania infantum. The enzyme catalyzes removal of the 14α-methyl group from sterol precursors. The reaction is essential for membrane biogenesis and therefore has great potential to become a target for antileishmanial chemotherapy. Although L. infantum CYP51 prefers C4-monomethylated sterol substrates such as C4-norlanosterol and obtusifoliol (V(max) of ∼10 and 8 min(-1), respectively), it is also found to 14α-demethylate C4-dimethylated lanosterol (V(max) = 0.9 min(-1)) and C4-desmethylated 14α-methylzymosterol (V(max) = 1.9 min(-1)). Binding parameters with six sterols were tested, with K(d) values ranging from 0.25 to 1.4 μM. Thus, L. infantum CYP51 is the first example of a plant-like sterol 14α-demethylase, where requirements toward the composition of the C4 atom substituents are not strict, indicative of possible branching in the postsqualene portion of sterol biosynthesis in the parasite. Comparative analysis of three CYP51 substrate binding cavities (Trypanosoma brucei, Trypanosoma cruzi, and L. infantum) suggests that substrate preferences of plant- and fungal-like protozoan CYP51s largely depend on the differences in the enzyme active site topology. These minor structural differences are also likely to underlie CYP51 catalytic rates and drug susceptibility and can be used to design potent and specific inhibitors.

PubMed: 21632531
DOI: 10.1074/jbc.M111.237099

Experimental method

X-RAY DIFFRACTION (2.75 Å)