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| Codon Optimization | The TSR1 domain of human ADAMTS5 corresponds to amino acid residues 567–622. The cDNA sequence corresponding to residues 561–634, which include the surrounding sequences on both the Nand C-terminus of TSR1, were codon-optimized for E. coli expression and cloned into a pET30a vector with a N-terminal His-tag (GenScript, Piscataway, NJ, USA). The membrane was blocked with 3% skimmed milk for 1 h at room temperature. This was followed by overnight incubation with an anti-His antibody (Cat # A00186-100, GenScript, Piscataway, NJ, USA), probing with the respective fluorescent tagged secondary antibody (Cat # 926-32212, LICOR Inc., Lincoln, NE, USA), and visualization using a fluorescent imaging system (Odyssey CLx, LI-COR Inc.). | Get A Quote |
Inhibiting tumor angiogenesis is a well-established approach for anticancer therapeutic development. A Disintegrin-like and Metalloproteinase with ThromboSpondin Motifs 5 (ADAMTS5) is a secreted matrix metalloproteinase in the ADAMTS family that also functions as an anti-angiogenic/anti-tumorigenic molecule. Its anti-angiogenic/anti-tumorigenic function is independent from its proteinase activity， but requires its first thrombospondin type 1 repeat (TSR1). However， it is not known if recombinant TSR1 (rTSR1) can function as an anticancer therapeutic. In this report， we expressed and purified a 75-residue recombinant TSR1 polypeptide from and investigated its ability to function as an anticancer therapeut... More
