ADAMTS-5, also known as implantin or aggrecanase-2, was first described as ADAMTS-5, a protein elevated in mice during the peri-implantion period. At the same time, another group identified aggrecanase-11, a protein elevated in arthritic synovium. The name was later changed to ADAMTS-5. ADAMTS-5 has been found in the heart, lung, cervix, uterus, ovary, brain, cartilage and numerous other tissues, as well as in chondroblastoma cell lines. Initial observations indicated a role for ADAMTS-5 in aggrecan cleavage and cartilage destruction, especially in arthritis, and potentially a role in embryo implantion. A member of the metalloproteinase family containing disintegrin-like domains (ADAMs) the function of ADAMTS-5 is still poorly understood. ADAMTS-5 contains the canonical HExxHxxxxxH zinc metalloproteinase motif, and has been shown to efficiently cleave aggrecan. In addition to the metalloprotease domain, ADAMTS-5 has a propeptide domain, a Prohormone Convertase (PC, furin) cleavage site, a cysteine-rich domain, a spacer domain and two thrombospondin-1 like domains. ADAMTS-5 is inhibited by the endogenous MMP inhibitors, TIMP-1, 2, 3 and 4 but most efficiently by TIMP-3. ADAMTS-5 does not have a transmembrane domain, unlike many of the ADAMs proteases, and is a secreted protein. Other ADAMs family members (ADAM-10, ADAM-17) have been more thoroughly studied, and are known to play key roles in inflammation, growth factor maturation and release, and a wide range of other functions. The full length ADAMTS-5 sequence codes for a 930 amino acid protein, with a predicted mass is 101.715 kD, but glycosylation and the abundance of cysteine residues gives ADAMTS-5 a greater apparent molecular weight on reduced SDS PAGE gels. ADAMTS-5 is cleaved at the furin cleavage site when it is secreted, with a predicted mass of 73.169 kD, which is further cleaved to generate a range of smaller forms. RP2ADAMTS-5 recognizes the zymogen of ADAMTS-5 at 120 kD in reduced Western blots, activated forms at 73 kD (major bands), and breakdown products at 50 kD and 40 kD in cell lysates. A recommended starting concentration for Western blots is 1:1000 when using colorimetric substrates such as BCIP/NBT, and 1:5000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species. EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen.

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