Abstract
In recent years, sea cucumber has become a favorite healthcare food due to its characteristic prevention of cardiovascular diseases, suppression of tumors, as well as enhancement of immunity. In order to screen the anti-tumoral proteins or peptides from sea cucumber (Apostichopus japonicus), its cDNA library was analyzed, and a disintegrin-like and metalloproteinase with thrombospondin type 1 motif, member 13 (ADAMTS13)-like was found. ADAMTS13-like contains 10 thrombospondin 1 (TSP1) domains. Based on analysis of bioinformatics, the third TSP1 domain of this protein, which is further named Aj-Tspin, contains an arginine-glycine-aspartate (RGD) motif. Since our previous studies showed that the recombinant RGD-containing peptide from lampreys showed anti-tumoral activity, the third TSP1 domain of ADAMTS13-like was chosen to evaluate it’s effect on tumor proliferation and metastasis, despite the fact it shares almost no homologue with disintegrins from other species. After artificial synthesis, its cDNA sequence, Aj-Tspin, which is composed of 56 amino acids, was subcloned into a pET23b vector and expressed as a recombinant Aj-Tspin (rAj-Tspin) in a soluble form with a molecular weight of 6.976 kDa. Through affinity chromatography, rAj-Tspin was purified as a single protein. Both anti-proliferation and immunofluorescence assays showed that rAj-Tspin suppressed the proliferation of Lewis lung carcinoma (LLC) cells through apoptosis. Adhesion assay also displayed that rAj-Tspin inhibited the adhesion of LLC cells to ECM proteins, including fibronectin, laminin, vitronectin and collagen. Lastly, rAj-Tspin also suppressed the migration and invasion of LLC cells across the filter in transwells. Thus, the above indicates that rAj-Tspin might act as a potential anti-tumoral drug in the future and could also provide information on the nutritional value of sea cucumber.
Highlights
A disintegrin-like and metalloproteinase with thrombospondin type 1 motif, member 13 (ADAMTS13) belongs to the ADAMTS family, which is composed of 19 proteases [1]
According to sequence analysis through the EXPASY website, ADAMTS13-like from A. japonicus is composed of 10 TSP1 domains (Figure 2)
Among the 10 TSP1 domains, only the third TSP1 domain was found to contain an RGD motif, which is the classic characterization of disintegrins (Figure A2)
Summary
A disintegrin-like and metalloproteinase with thrombospondin type 1 motif, member 13 (ADAMTS13) belongs to the ADAMTS family, which is composed of 19 proteases [1]. Except the domains shared by the 19 ADAMTS members, human ADAMTS13 has eight thrombospondin type 1 motifs and has been most reported to affect the progress of thrombotic thrombocytopenic purpura [1,2]. During the past 20 years, identification of the biological characteristics of NSCLC and its treatment strategies have progressed to a significant degree. Both targeted therapy and immunotherapy have provided novel information for treatment with controllable toxicity, which could prolong the lifespan of NSCLC patients. More effective drugs targeted for NSCLC patients are still required to be screened and verified
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
