ADAMTS-9 is a member of the ADAMs family of proteinases with Thrombospondin motifs. ADAMTS-9 is closest in homology to ADAMTS-20, sharing 54% overall identity, and like ADAMTS-20 ADAMTS-9 gas a GON-like domain at the carboxyterminal end. The GON domain at the carboxyterminal end is similar to the GON-1 protein in C. elegans, mutations of which lead to defective gonadal development. ADAMTS-9 is expressed in ovary and testis, but little is known about the role of ADAMTS-9 in reproductive organs. ADAMTS-9 is also expressed in the heart, placenta, lung, skeletal tissue, and pancreas, so the protein must have wider functions than just gonadal development. ADAMTS-9, like ADAMTS-20, has a total of 15 thrombospondin-like domains. The first TS domain begins shortly after the catalytic and disintegrin domains. TS domains 2-6 follow a spacer domain, followed by linker domain 1, then TS domains 7 & 8, linker domain 2, and then TS domains 9-15. In other ADAMTS proteins the TS motifs are thought to bind to the ECM. The ADAMTS proteins contain at least one prohormone-convertase cleavage site, although it appears that one site is used preferentially, and cleavage of the propeptide domain at this site generates active enzymes. For ADAMTS-9, this site is the RRTKR sequence, and the mature ADAMTS-9 has an aminoterminal sequence of FLSYPR. The catalytic site of ADAMTS-9 is most like ADAMTS-1, 14 and 15, with an HExxHVFNMxH sequence, perhaps giving these enzymes some shared specificity. Several sequences have been found for human ADAMTS-9, a 1935 amino acid sequence with predicted mass of 216.5 kDa and a pI of 7.464, a 1629 amino acid sequence with predicted mass of 182.65 and pI of 7.36, and a 1072 amino acid sequence with predicted mass of 120.65 kDa and pI of 7.618. The sequences are identical up to the TS5 repeat, where the shortest sequence ends. The second sequence ends in the TS14 repeat, and only the longest sequence contains the GON-like domain. It is not clear if the different sequence lengths are due to alternative slicing of the gene or incomplete sequencing. A recommended starting concentration for Western blots is 1:1,000 when using colorimetric substrates such as BCIP/NBT, and 1:5,000 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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