The ADAM10 prodomain is a specific inhibitor of ADAM10 proteolytic activity and inhibits cellular shedding events.
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-alpha-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.
Moss
Marcia L
ML
BioZyme Incorporated, Apex, North Carolina 27523, USA. mmoss@biozyme-inc.com
Bomar
Martha
M
Liu
Qian
Q
Sage
Harvey
H
Dempsey
Peter
P
Lenhart
Patricia M
PM
Gillispie
Patricia A
PA
Stoeck
Alexander
A
Wildeboer
Dirk
D
Bartsch
Jörg W
JW
Palmisano
Ralf
R
Zhou
Pei
P
D051722:ADAM Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism* / Q000627:therapeutic use; D000072197:ADAM10 Protein; D000072198:ADAM17 Protein; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors* / Q000378:metabolism* / Q000627:therapeutic use; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D066259:Betacellulin; D019556:COS Cells; D002522:Chlorocebus aethiops; D019483:Disintegrins / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000627:therapeutic use; D004792:Enzyme Precursors / Q000037:antagonists & inhibitors* / Q000378:metabolism* / Q000627:therapeutic use; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism* / Q000627:therapeutic use; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy / Q000201:enzymology; D017434:Protein Structure, Tertiary / Q000502:physiology
D016564:Amyloid beta-Protein Precursor; D015703:Antigens, CD; C587851:BTC protein, human; D066259:Betacellulin; C587852:Btc protein, mouse; D019483:Disintegrins; D004792:Enzyme Precursors; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C512478:EGFR protein, human; D066246:ErbB Receptors; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; C489402:Adam8 protein, mouse; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human; C000606463:Adam17 protein, mouse