Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - A disintegrin and metalloproteinase (ADAM)-mediated ectodomain shedding of ADAM10
AU - Parkin, Edward
AU - Harris, Benjamin
PY - 2009/3
Y1 - 2009/3
N2 - A disintegrin and metalloproteinase (ADAM) 10 is a type I transmembrane glycoprotein responsible for the ectodomain shedding of a range of proteins including the amyloid precursor protein implicated in Alzheimer's disease. In this study we demonstrate that ADAM10 itself is subject to shedding by one or more ADAMs. Expression of epitope-tagged wild-type ADAM10 in SH-SY5Y cells enabled the detection of a soluble ectodomain in conditioned medium. Shedding of the ADAM10 ectodomain was inhibited by a known ADAM inhibitor with a reciprocal accumulation of the full-length mature protein in both cell lysates and extracellular membrane vesicles. Shedding was also stimulated by phorbol ester treatment of cells. A glycosylphosphatidylinositol-anchored form of ADAM10 lacking the cytosolic, transmembrane and alpha-helical juxtamembrane regions of the wild-type protein was shed in a similar manner. Furthermore, a truncated soluble ADAM10 construct, although correctly post-translationally processed and catalytically active against a synthetic peptide substrate, was incapable of shedding cell-associated amyloid precursor protein. Finally, we show that ADAM9 is, at least in part, responsible for the ectodomain shedding of ADAM10. In conclusion, this is a new mechanism by which levels of ADAM10 are regulated and may have implications in a range of human diseases including Alzheimer's disease.
AB - A disintegrin and metalloproteinase (ADAM) 10 is a type I transmembrane glycoprotein responsible for the ectodomain shedding of a range of proteins including the amyloid precursor protein implicated in Alzheimer's disease. In this study we demonstrate that ADAM10 itself is subject to shedding by one or more ADAMs. Expression of epitope-tagged wild-type ADAM10 in SH-SY5Y cells enabled the detection of a soluble ectodomain in conditioned medium. Shedding of the ADAM10 ectodomain was inhibited by a known ADAM inhibitor with a reciprocal accumulation of the full-length mature protein in both cell lysates and extracellular membrane vesicles. Shedding was also stimulated by phorbol ester treatment of cells. A glycosylphosphatidylinositol-anchored form of ADAM10 lacking the cytosolic, transmembrane and alpha-helical juxtamembrane regions of the wild-type protein was shed in a similar manner. Furthermore, a truncated soluble ADAM10 construct, although correctly post-translationally processed and catalytically active against a synthetic peptide substrate, was incapable of shedding cell-associated amyloid precursor protein. Finally, we show that ADAM9 is, at least in part, responsible for the ectodomain shedding of ADAM10. In conclusion, this is a new mechanism by which levels of ADAM10 are regulated and may have implications in a range of human diseases including Alzheimer's disease.
KW - ADAM Proteins
KW - Amyloid Precursor Protein Secretases
KW - Amyloid beta-Protein Precursor
KW - Arginine
KW - Cell Line
KW - Disintegrins
KW - Enzyme Inhibitors
KW - Humans
KW - Membrane Proteins
KW - Metalloproteases
KW - Mutation
KW - Neuroblastoma
KW - Peptides
KW - Phosphoinositide Phospholipase C
KW - Protein Structure, Tertiary
KW - Protein Transport
KW - RNA, Small Interfering
KW - Tetradecanoylphorbol Acetate
KW - Transfection
UR - http://www.scopus.com/inward/record.url?scp=61349132079&partnerID=8YFLogxK
U2 - 10.1111/j.1471-4159.2009.05907.x
DO - 10.1111/j.1471-4159.2009.05907.x
M3 - Journal article
C2 - 19183255
VL - 108
SP - 1464
EP - 1479
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
SN - 1471-4159
IS - 6
ER -