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Neurotropic and neuroprotective activities of the earthworm peptide Lumbricusin
Authors:Dae Hong Kim  Ik Hwan Lee  Seung Taek Nam  Ji Hong  Peng Zhang  Jae Sam Hwang  Heon Seok  Hyemin Choi  Dong Gun Lee  Jae Il Kim  Ho Kim
Affiliation:1. Department of Life Science, College of Natural Science, Daejin University, Pocheon, Gyeonggido 487-711, South Korea;2. Department of Agricultural Biology, National Academy of Agricultural Science, RDA, Suwon 441-707, South Korea;3. Department of Biomedical Engineering, Jungwon University, Goesan, Chungcheongbukdo 367-700, South Korea;4. School of Life Sciences, KNU Creative Bioresearch Group (BK21 Plus Program), College of Natural Sciences, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 702-701, South Korea;5. School of Life Sciences, Gwangju Institute of Science and Technology, Oryong-dong, Buk-gu, Gwangju 500-712, South Korea
Abstract:We recently isolated a polypeptide from the earthworm Lumbricus terrestris that is structurally similar to defensin, a well-known antibacterial peptide. An 11-mer antibacterial peptide (NH2-RNRRWCIDQQA), designated Lumbricusin, was synthesized based on the amino acid sequence of the isolated polypeptide. Since we previously reported that CopA3, a dung beetle peptide, enhanced neuronal cell proliferation, we here examined whether Lumbricusin exerted neurotropic and/or neuroprotective effects. Lumbricusin treatment induced a time-dependent increase (∼51%) in the proliferation of human neuroblastoma SH-SY5Y cells. Lumbricusin also significantly inhibited the apoptosis and decreased viability induced by treatment with 6-hydroxy dopamine, a Parkinson’s disease-mimicking agent. Immunoblot analyses revealed that Lumbricusin treatment increased ubiquitination of p27Kip1 protein, a negative regulator of cell-cycle progression, in SH-SY5Y cells, and markedly promoted its degradation. Notably, adenoviral-mediated over-expression of p27Kip1 significantly blocked the antiapoptotic effect of Lumbricusin in 6-hydroxy dopamine-treated SH-SY5Y cells. These results suggest that promotion of p27Kip1 degradation may be the main mechanism underlying the neuroprotective and neurotropic effects of Lumbricusin.
Keywords:SH-SY5Y, neuroblastoma cells   6-OHDA, 6-hydroxy dopamine   MTT, 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide   FACS, fluorescence-activated cell sorter   DMSO, dimethyl sulfoxide   p21, p27Kip1   IP, immunoprecipitation   GFP, green fluorescent protein
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