1. Discovering active sites in peptide Ala-Val-Thr-Phe that counter 2,2-azobis(2-methylpropanimidamidine)dihydrochloride-induced oxidative stress in HepG2 cells
Jiaxi Liang, Qin Wang, Jianliang Liu, Guozhong Huang, Churong Liang, Huifan Liu, Lukai Ma RSC Adv. 2020 Jun 26;10(41):24444-24453. doi: 10.1039/d0ra02292f. eCollection 2020 Jun 24.
The Ala-Val-Thr-Phe (AVTF) peptide derived from edible Dendrobium aphyllum was co-incubated with Lactobacillus amylolyticus in a previous study. The aim of the present study was to further examine the antioxidative and protective effects of the AVTF peptides through the analysis of free-radical quenching in HepG2 cells subjected to 2,2-azobis(2-methylpropanimidamidine)dihydrochloride (AAPH)-induced oxidative stress and to determine the active sites within the peptide. Variations in intracellular malondialdehyde levels indicated that these peptides protect HepG2 cells by preventing ROS attack and lipid peroxidation. Antioxidant enzymes and Nrf2 were downregulated in AVTF-treated but not in AAPH-treated HepG2 cells, whereas the electrically sensitive Keap1 was not susceptible to free radical-induced damage after AVTF treatment. However, this did not result in the activation of the Nrf2/Keap1 signaling pathway, thus indicating that one potential mechanism by which AVTF maintains homeostasis in HepG2 cells is by directly scavenging free radicals. Furthermore, quantum chemical calculations and the assessment of electronic-related properties associated with antioxidant activity revealed that the active sites of AVTF included N9-H11, which was further confirmed by the assessment of ROS levels in methylated AVTF-treated cells. The results of this study provide valuable insights into the active site N9-H11 in the Ala residue of AVTF, which influences the antioxidant activity of the peptide.
2. Total Synthesis of Alloviroidin
Carol M Taylor, Samuel K Kutty, Benson J Edagwa Org Lett. 2019 Apr 5;21(7):2281-2284. doi: 10.1021/acs.orglett.9b00567. Epub 2019 Mar 12.
Alloviroidin is a cyclic heptapeptide, produced by several species of Amanita mushrooms, that demonstrates high affinity for F-actin as is characteristic of virotoxins and phallotoxins. Alloviroidin was synthesized via a [3 + 4] fragment condensation of Fmoc-d-Thr(OTBS)-d-Ser(OTBS)-(2 S,3 R,4 R)-DHPro(OTBS)2-OH and H-Ala-Trp(2-SO2Me)-(2 S,4 S)-DHLeu(5-OTBS)-Val-OMe to form bond A. The linear heptapeptide favored a turn conformation, facilitating cyclization between Val1 and d-Thr2 (position B). Global deprotection and HPLC purification afforded alloviroidin with NMR spectra in excellent agreement with the natural product.
3. Oostatic peptides
Jan Hlaváček Amino Acids. 2013 Apr;44(4):1095-105. doi: 10.1007/s00726-012-1449-x. Epub 2013 Jan 8.
Oostatic peptides are organic molecules, which influence an insect reproduction due to a regulation of the eggs development. It was proved that decapeptide-H-Tyr-Asp-Pro-Ala-Pro-Pro-Pro-Pro-Pro-Pro-OH (YDPAPPPPPP)-isolated from mosquito Aedes aegypti, inhibits trypsin activity in the midgut of the mosquito. Therefore, it was named trypsin-modulating oostatic factor (Aea-TMOF). Feeding the recombinant cells with cloned and expressed TMOF on the coat protein of tobacco mosaic virus (TMV) to mosquito larvae, caused larval mortality. The TMOF was therefore designed for usage as a new biorational insecticide against mosquito. Similarly, a hexapeptide-H-Asn-Pro-Thr-Asn-Leu-His-OH (NPTNLH)-was isolated from the grey flesh fly Neobellieria bullata. This peptide and some of its analogs inhibited trypsin-like synthesis by the midgut in female flies and was therefore entitled Neb-TMOF. Interestingly, the synthetic Aea-TMOF and mainly its C-terminus shorten analogs, including those containing D-amino acids or methylene-oxy isosteric bond, quickly and strongly inhibited the hatchability and egg development in the flesh fly N. bullata.
