Synthesis of lucifensin by native chemical ligation and characteristics of its isomer having different disulfide bridge pattern

0431796 - ÚOCHB 2015 RIV GB eng J - Journal Article
Stanchev, Stancho - Zawada, Zbigniew - Monincová, Lenka - Bednárová, Lucie - Slaninová, Jiřina - Fučík, Vladimír - Čeřovský, Václav
Synthesis of lucifensin by native chemical ligation and characteristics of its isomer having different disulfide bridge pattern.
Journal of Peptide Science. Roč. 20, č. 9 (2014), s. 725-735. ISSN 1075-2617. E-ISSN 1099-1387
Institutional support: RVO:61388963
Keywords : lucifensin * native chemical ligation * solid-phase peptide synthesis * antimicrobial activity * CD spectroscopy * DTT reduction
Subject RIV: CE - Biochemistry
Impact factor: 1.546, year: 2014

The antimicrobial 40-amino-acid-peptide lucifensin was synthesized by native chemical ligation (NCL) using N-acylbenzimidazolinone (Nbz) as a linker group. NCL is a method in which a peptide bond between two discreet peptide chains is created. Thismethod has been applied to the synthesis of long peptides and proteins when solid-phase synthesis is imcompatible. Two models of ligation were developed: [15 + 25] Ala-Cys and [19 + 21] His-Cys. The [19 + 21] His-Cys method gives lower yield because of the lower stability of 18-peptide-His-Nbz-CONH2 peptide, as suggested by density functional theory calculation. Acetamidomethyldeprotection and subsequent oxidation of the ligated linear lucifensin gave a mixture of lucifensin isomers, which differed in the location of their disulfide bridges only. The dominant isomer showed unnatural pairing of cysteines [C1 - 6], [C3 - 5], and [C2 - 4], which limits its ability to form alpha-helical structure. The activity of isomeric lucifensin toward Bacillus subtilis, Staphylococcus aureus, and Micrococcus luteus was lower than that of the natural lucifensin. The desired product native lucifensin was prepared from this isomer using a one-pot reduction with dithiotreitol and subsequent air oxidation in slightly alkaline medium.
Permanent Link: http://hdl.handle.net/11104/0236422