Dimensional mapping of the active site of cholesterol esterase with alkylboronic acid inhibitors.
The cholesterol esterase-catalyzed hydrolysis of the water-soluble substrate p-nitrophenyl butyrate occurs via an acylenzyme mechanism, and is competitively inhibited by boronic acid transition state analog inhibitors. Accordingly, we undertook to dimensionally map the enzyme's active site via synthesis and characterization of a series of n-alkyl boronic acid inhibitors. The most potent of these is n-hexaneboronic acid, with a Ki = 13 +/- 1 microM, since inhibitor potency declines for both longer and shorter boronic acids. No inhibition is observed for methaneboronic acid and n-octaneboronic acid inhibits poorly, with a Ki of 7 mM. These results indicate that the ability of the enzyme to form tight complexes with boron-containing transition state analog inhibitors is sensitive to alkyl chain length. The trend in inhibitor potency is discussed in terms of substrate specificity of and transition state stabilization by cholesterol esterase, and has important implications for the design of optimal reversible inhibitors of the enzyme.
Sutton
L D
LD
Department of Chemistry, University of Iowa, Iowa City 52242.
Lantz
J L
JL
Eibes
T
T
Quinn
D M
DM
D000478:Alkylation; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D001897:Boronic Acids / Q000494:pharmacology*; D010179:Pancreas / Q000201:enzymology; D002787:Sterol Esterase / Q000037:antagonists & inhibitors*; D013329:Structure-Activity Relationship; D013552:Swine
D001897:Boronic Acids; D002787:Sterol Esterase