Abstract
In the venom of eusocial bee Lasioglossum laticeps, we identified a novel unique antimicrobial peptide named lasiocepsin consisting of 27 amino acid residues and two disulfide bridges. After identifying its primary structure, we synthesized lasiocepsin by solid-phase peptide synthesis using two different approaches for oxidative folding. The oxidative folding of fully deprotected linear peptide resulted in a mixture of three products differing in the pattern of disulfide bridges. Regioselective disulfide bond formation significantly improved the yield of desired product. The synthetic lasiocepsin possessed antimicrobial activity against both Gram-positive and -negative bacteria, antifungal activity against Candida albicans, and no hemolytic activity against human erythrocytes. We synthesized two lasiocepsin analogs cyclized through one native disulfide bridge in different positions and having the remaining two cysteines substituted by alanines. The analog cyclized through a Cys8–Cys25 disulfide bridge showed reduced antimicrobial activity compared to the native peptide while the second one (Cys17–Cys27) was almost inactive. Linear lasiocepsin having all four cysteine residues substituted by alanines or alkylated was also inactive. That was in contrast to the linear lasiocepsin with all four cysteine residues non-paired, which exhibited remarkable antimicrobial activity. The shortening of lasiocepsin by several amino acid residues either from the N- or C-terminal resulted in significant loss of antimicrobial activity. Study of Bacillus subtilis cells treated by lasiocepsin using transmission electron microscopy showed leakage of bacterial content mainly from the holes localized at the ends of the bacterial cells.
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Acknowledgments
This work was supported by the Czech Science Foundation, Grants Nos. 203/08/0536 and P205/10/1276, and by Research Project No. Z40550506 of the Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic. We thank our technical assistant Mrs. Hana Hulačová for the help with peptide synthesis. Lenka Monincová thanks the Ministry of Education of the Czech Republic for a stipend and for additional financial support from Specific University Research Project No. 33779266 awarded by Charles University Prague. We also thank Gale A. Kirking at English Editorial Services, s.r.o. for assistance with the English.
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Monincová, L., Slaninová, J., Fučík, V. et al. Lasiocepsin, a novel cyclic antimicrobial peptide from the venom of eusocial bee Lasioglossum laticeps (Hymenoptera: Halictidae). Amino Acids 43, 751–761 (2012). https://doi.org/10.1007/s00726-011-1125-6
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DOI: https://doi.org/10.1007/s00726-011-1125-6