The ADAM10 Prodomain Is a Specific Inhibitor of ADAM10 Proteolytic Activity and Inhibits Cellular Shedding Events

Abstract

ADAM10 is a disintegrin metalloproteinase that processes amyloid precursor protein and ErbB ligands and is involved in the shedding of many type I and type II single membrane-spanning proteins. Like tumor necrosis factor-␣-converting enzyme (TACE or ADAM17), ADAM10 is expressed as a zymogen, and removal of the prodomain results in its activation. Here we report that the recombinant mouse ADAM10 prodomain, purified from Escherichia coli, is a potent competitive inhibitor of the human ADAM10 catalytic/disintegrin domain, with a K i of 48 nM. Moreover, the mouse ADAM10 prodomain is a selective inhibitor as it only weakly inhibits other ADAM family proteinases in the micromolar range and does not inhibit members of the matrix metalloproteinase family under similar conditions. Mouse prodomains of TACE and ADAM8 do not inhibit their respective enzymes, indicating that ADAM10 inhibition by its prodomain is unique. In cell-based assays we show that the ADAM10 prodomain inhibits betacellulin shedding, demonstrating that it could be of potential use as a therapeutic agent to treat cancer.

Shedding of Endogenous Interleukin-6 Receptor (IL-6R) Is Governed by A Disintegrin and Metalloproteinase (ADAM) Proteases while a Full-length IL-6R Isoform Localizes to Circulating Microvesicles

“…This strongly suggests that ADAM10 is also responsible for the constitutive release of endogenous IL-6R in the mouse. We also assessed constitutive IL-6R shedding in the presence of recombinant murine ADAM10 prodomain, which has been reported to effectively block ADAM10 activity in the mouse (20). Supporting our results with pharmacological inhibitors we also noticed a substantial reduction of constantly released IL-6R when murine ADAM10 was blocked by the inhibitory prodomain (Fig.…”

“…Cells were stimulated with PMA or LPS from Escherichia coli O111:B4 (both Sigma) as depicted in the figure legends. Generation of neutralizing, bivalent ADAM17 antibody D1(A12) and recombinant murine ADAM10 prodomain were described previously (19,20). The hydroxamate-based ADAM inhibitors GI254023X and GW280264X were synthesized by Iris Biotech (Marktredwitz, Germany).…”

Translational Repression of the Disintegrin and Metalloprotease ADAM10 by a Stable G-quadruplex Secondary Structure in Its 5′-Untranslated Region

“…ADAM10 is expressed as an inactive prodomain containing zymogene, which is activated by the proprotein convertases furin and PC7 (14). The prodomain is important for folding of ADAM10 and acts as a potent inhibitor of ADAM10 activity (14,15). Recently, it was demonstrated that ADAM10 transcription is stimulated by all-trans retinoic acid and by the deacetylase Sirtuin1 (16,17).…”

Sequential and γ-secretase-dependent processing of the betacellulin precursor generates a palmitoylated intracellular-domain fragment that inhibits cell growth

“…S1A) was capable of ADAM10-dependent BTC shedding and the generation of a stable, cellular BTC remnant (termed BTC-CTF) (Moss et al, 2007). To investigate the biological significance of the BTC-CTF in IMPE cells expressing BTC-WT (IMPE-BTC-WT), we first verified that proBTC was correctly processed to produce BTC-CTF (Moss et al, 2007;Sanderson et al, 2006a;Sanderson et al, 2005).…”

“…We have previously demonstrated that the extracellular domain of proBTC can be proteolytically cleaved by ADAM10 to release the mature, active ligand and to produce a stable, cellular BTC remnant (termed BTC-CTF) (Moss et al, 2007;Sanderson et al, 2006a;Sanderson et al, 2005). Because of its stability, BTC-CTF represents a significant proportion of the cellular BTC isoforms expressed in cells under steady-state conditions.…”