Metalloprotease inhibitor profiles of human ADAM8 in vitro and in cell-based assays.

Metalloprotease inhibitor profiles of human ADAM8 in vitro and in cell-based assays. Biol Chem. 2018 Dec 01;: Authors: Schlomann U, Dorzweiler K, Nuti E, Tuccinardi T, Rossello A, Bartsch JW Abstract ADAM8 as a membrane-anchored metalloproteinase-disintegrin is upregulated under pathological conditions such as inflammation and cancer. As active sheddase, ADAM8 can cleave several membrane proteins, among them the low-affinity receptor FcεRII CD23. Hydroxamate-based inhibitors are routinely used to define relevant proteinases involved in ectodomain shedding of membrane proteins. However, for ADAM proteinases, common hydroxamates have variable profiles in their inhibition properties, commonly known for ADAM proteinases 9,10, and 17. Here we determined the inhibitor profile of human ADAM8 for eight ADAM/MMP inhibitors by in vitro assays using recombinant ADAM8 as well as the in vivo inhibition in cell-based assays using HEK293 cells to monitor release of soluble CD23 by ADAM8. ADAM8 activity is inhibited by BB94 (batimastat), GW280264, FC387 and FC143 (two ADAM17 inhibitors), weaker by GM6001, TAPI2 and BB2516 (marimastat), while no inhibition was observed for GI254023, an ADAM10 specific inhibitor. Modelling of inhibitor FC143 bound to the catalytic sites of ADAM8 and ADAM17 reveals similar geometries in the pharmacophoric regions of both proteinases, which is different in ADAM10 due to replacement in the S1 position of T300 (ADAM8) an...
Source: Biological Chemistry - Category: Chemistry Tags: Biol Chem Source Type: research