Fibrinogen-related proteins play important roles in the immune responses. We have obtained a cDNA encoding a novel fibrinogen-related protein from roughskin sculpin Trachidermus fasciatus (T. fasciatus) and named it as TfFREP2. The N and C terminus of TfFREP2 contain a putative 21-amino acid signal peptide and a typical 217-amino acid fibrinogen-like domain, which is conserved in all fibrinogen-related proteins. TfFREP2 has three glycosylation sites and two potential calcium-binding sites that are possibly involved in calcium coordination. The results of tissue specific checking showed that the mRNA and protein of TfFREP2 were particularly abundant in skin and gill among all the tested tissues. TfFREP2 mRNA and protein expression changed significantly after being challenged by V. anguillarum pathogen in those immune-barrier tissues, such as skin and gill. Furthermore, recombinant TfFREP2 is able to agglutinate and bind V. anguillarum in the presence of calcium ion. The above results suggest that TfFREP2 might be involved in the host defense of fish against V. anguillarum infection.