Identification of an LPS-Induced Chemo-Attractive Peptide from Ciona robusta

Background: Previously published work has demonstrated that the LPS injection ofCiona robusta leads to the overexpression of a truncated form of an immune-related mRNA (C8short)by means of Ciona robusta (CR) alternative polyadenylation (APA) (CR-APA). Methods: The 3Dstructure of the C8short-derived Ciona robusta chemo-attractive peptide (CrCP) was evaluated byhomology modeling. The biological activity of the CrCP was studied in vitro using a primary humandermal cell line (HuDe). Real-Time PCR was used to investigate the expression levels of genesinvolved in cell motility. NF-?B signaling was studied by western blotting. Results: In silico modelingshowed that CrCP displayed structural characteristics already reported for a short domain of thevertebrate CRK gene, suggesting its possible involvement in cell migration mechanisms. In vitroassays demonstrated that CrCP was capable of inducing the motility of HuDe cells in both woundhealing and chemo-attractive experiments. qPCR demonstrated the capability of CrCP to modulatethe expression of the matrix metalloproteinase-7 (MMP-7) and E-cadherin genes. Finally, western blotanalysis demonstrated that treatment with CrCP induced activation of the NF-?B signaling pathway.Conclusion: Our results describe the characterization of the 3D structure and chemo-attractive activityof an LPS-induced CrCP peptide from Ciona robusta.