Lecithin:cholesterol acyltransferase activation by sulfhydryl-reactive small molecules: Role of cysteine-31
Articolo
Data di Pubblicazione:
2017
Citazione:
Lecithin:cholesterol acyltransferase activation by sulfhydryl-reactive small molecules: Role of cysteine-31 / L.A. Freeman, S. J. Demosky Jr., M. Konaklieva, R. Kuskovsky, A. Aponte, A.F. Ossoli, S.M. Gordon, R.F. Koby, K.A. Manthei, M. Shen, B.L. Vaisman, R.D. Shamburek, A. Jadhav, L. Calabresi, M. Gucek, J.J.G. Tesmer, R.L. Levine, A.T. Remaley. - In: THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS. - ISSN 0022-3565. - 362:2(2017 Aug 01), pp. 306-318. [10.1124/jpet.117.240457]
Abstract:
Lecithin: cholesterol acyltransferase (LCAT) catalyzes plasma cholesteryl ester formation and is defective in familial lecithin: cholesterol acyltransferase deficiency (FLD), an autosomal recessive disorder characterized by low high-density lipoprotein, anemia, and renal disease. This study aimed to investigate the mechanism by which compound A [3-(5-(ethylthio)-1,3,4-thiadiazol-2-ylthio)pyrazine-2-carbonitrile], a small heterocyclic amine, activates LCAT. The effect of compound A on LCAT was tested in human plasma and with recombinant LCAT. Mass spectrometry and nuclear magnetic resonance were used to determine compound A adduct formation with LCAT. Molecular modeling was performed to gain insight into the effects of compound A on LCAT structure and activity. Compound A increased LCAT activity in a subset (three of nine) of LCAT mutations to levels comparable to FLD heterozygotes. The site-directed mutation LCAT-Cys31 Gly prevented activation by compound A. Substitution of Cys31 with charged residues (Glu, Arg, and Lys) decreased LCAT activity, whereas bulky hydrophobic groups (Trp, Leu, Phe, and Met) increased activity up to 3-fold (P, < 0.005). Mass spectrometry of a tryptic digestion of LCAT incubated with compound A revealed a + 103.017 m/z adduct on Cys31, consistent with the addition of a single hydrophobic cyanopyrazine ring. Molecular modeling identified potential interactions of compound A near Cys31 and structural changes correlating with enhanced activity. Functional groups important for LCAT activation by compound A were identified by testing compound A derivatives. Finally, sulfhydryl-reactive beta-lactams were developed as a new class of LCAT activators. In conclusion, compound A activates LCAT, including some FLD mutations, by forming a hydrophobic adduct with Cys31, thus providing a mechanistic rationale for the design of future LCAT activators.
Tipologia IRIS:
01 - Articolo su periodico
Elenco autori:
L.A. Freeman, S. J. Demosky Jr., M. Konaklieva, R. Kuskovsky, A. Aponte, A.F. Ossoli, S.M. Gordon, R.F. Koby, K.A. Manthei, M. Shen, B.L. Vaisman, R.D. Shamburek, A. Jadhav, L. Calabresi, M. Gucek, J.J.G. Tesmer, R.L. Levine, A.T. Remaley
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